Anti-sortilin antibodies and methods of use thereof

ABSTRACT

The present disclosure is generally directed to compositions that include antibodies, e.g., monoclonal, chimeric, affinity-matured, humanized antibodies, antibody fragments, etc., that specifically bind a Sortilin protein, e.g., human Sortilin or mammalian Sortilin, and have improved and/or enhanced functional characteristics, and use of such compositions in preventing, reducing risk, or treating an individual in need thereof.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.62/698,007, filed Jul. 13, 2018, U.S. Provisional Application No.62/860,184, filed Jun. 11, 2019, and U.S. Provisional Application No.62/868,849, filed Jun. 28, 2019, each of which is hereby incorporated byreference in its entirety.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

This invention was made with government support under Award NumberR44AG050363 awarded by the National Institutes of Health. The governmenthas certain rights in the invention.

SUBMISSION OF SEQUENCE LISTING ON ASCII TEXT FILE

The content of the following submission on ASCII text file isincorporated herein by reference in its entirety: a computer readableform (CRF) of the Sequence Listing (file name: 735022001900SEQLIST.TXT,date recorded: Jul. 10, 2019, size: 153 KB).

FIELD

This present disclosure relates to anti-Sortilin antibodies, andtherapeutic uses of such antibodies.

BACKGROUND

Sortilin is a Type I transmembrane protein that acts both as a receptorof several ligands, and in the sorting of select cargo from thetrans-Golgi network (TGN) to late endosomes and lysosomes fordegradation. Sortilin harbors a large extracellular domain that is partof the VPS10 family, homologous to yeast VPS101P, and contains of a10-blade beta-propeller structure and a cysteine-rich 10CC module(Nykjaer, A et al., (2012) Trends Neurosci 35: 261-270; and Zheng, Y etal., (2011) PLoS One 6: e21023). A small fraction of Sortilin may beshed by activity of ADAM10 or gamma-Secretase (<5%) (Nykjaer, A et al.,(2012) Trends Neurosci 35: 261-270; and Willnow, T E et al., (2011) CurrOpin Lipidol 22: 79-85).

Sortilin binds the secreted protein Progranulin (PGRN) and targets itfor lysosomal degradation, thus negatively regulating extracellularlevels of PGRN (Hu, F et al. (2010) Neuron 68, 654-667. In line withthis, deficiency of Sortilin significantly increases plasma PGRN levelsboth in mouse models in vivo and human cells in vitro (Carrasquillo, M.M et al., (2010) Am J Hum Genet 87, 890-897; Lee, W. C et al., (2014)Hum Mol Genet 23, 1467-1478). Moreover, a polymorphism in Sortilin wasshown to be strongly assoiated with PGRN serum levels in humans(Carrasquillo M M et al., (2010), Am J Hum Genet. 10; 87(6):890-7).

Progranulin (PGRN) is a secreted, growth factor-like, trophic, andanti-inflammatory protein, which also plays a role as an adipokineinvolved in diet-induced obesity and insulin resistance (Nguyen D A etal., (2013). Trends in Endocrinology and Metabolism, 24, 597-606).Progranulin deficiency accounts for roughly 25% of all heritable formsof frontotemporal dementia (FTD), an early-onset neurodegenerativedisease. Patients with heterozygous loss-of-function mutations in PGRNhave ˜50% reduced extracellular levels of the protein and they willinvariably develop FTD, making PGRN a causal gene for the disease(Baker, M et al., (2006) Nature 442, 916-919; Carecchio M et al., (2011)J Alzheimers Dis 27, 781-790; Cruts, M et al., (2008) Trends Genet 24,186-194; Galimberti, D et al., (2010) J Alzheimers Dis 19, 171-177). Inaddition, PGRN mutant alleles have been identified in Alzheimer'sdisease patients (Seelaar, H et al., (2011). Journal of neurology,neurosurgery, and psychiatry 82, 476-486). Importantly, PGRN actsprotectively in several disease models with increased PGRN levels,accelerating behavioral recovery from ischemia (Tao, J et al., (2012)Brain Res 1436, 130-136; Egashira, Y. et al., (2013). JNeuroinflammation 10, 105), suppressing locomotor deficits in aParkinson's disease model (Van Kampen, J. M et al. (2014). PLoS One 9,e97032), attenuating pathology in a model of amyotriphic lateralsclerosis (Laird, A. S et al., (2010). PLoS One 5, e13368.) andarthritis (Tang, W et al., (2011). Science 332, 478-484) and preventingmemory deficits in an Alzheimer's disease model (Minami, S. S et al.,(2014). Nat Med 20, 1157-1164).

Sortilin also binds directly to pro-neurotrophins, such as pro-nervegrowth factor (pro-NGF), pro-BDNF, pro-neurotrophin-3, etc., whichharbor a pro-domain and are typically pro-apoptotic. Suchpro-neurotrophin precursors are released during stress, and Sortilin isinvolved in regulating their release as well as binding on the receivingcell and stimulation of apoptosis in conjunction with p75NTR (Willnow, TE et al., (2008) Nat Rev Neurosci 9: 899-909; Nykjaer, A et al., TrendsNeurosci 35: 261-270; and Nykjaer, A et al., (2004) Nature 427: 843-848;Hiroko Yano et al., (2009) J Neurosci.; 29: 14790-14802. Teng H. K., etal., J. Neurosci. 25:5455-5463 (2005)). Sortilin also binds to p75NTRdirectly (Skeldal S et al., (2012) J Biol Chem.; 287:43798). Sortilinalso binds to neurotensin in a region that partially overlaps withProgranulin binding (Quistgaard, E M et al., (2009) Nat Struct Mol Biol16: 96-98; and Zheng, Y et al., PLoS One 6: e21023). Sortilin alsointeracts with the Trk receptors NTRK1, NTRK2, and NTRK3; and canregulate their anterograde axonal transport and signaling (Vaegter, C Bet al., (2011) Nat. Neurosci. 14:54-61). Sortilin also interacts withand regulates the processing and trafficking of amyloid precursorprotein and the resulting production of pathological beta amyloidpeptides (Gustafsen C et al., (2013). J Neurosci. 2; 33 (1):64-71).

Sortilin has also been shown to bind to apolipoproteins and lipoproteinlipase; thus, deficiency leads to reduced VLDL release from liver andreduced cholesterol (Willnow, T E et al., (2011) Curr Opin Lipidol 22:79-85; Kjolby, M et al., (2010) Cell Metab 12: 213-223; Nilsson, S K etal., (2007) Biochemistry 46: 3896-3904.; Nilsson, S K et al., (2008) JBiol Chem 283: 25920-25927; and Klinger, S C et al., (2011)J Cell Sci124: 1095-1105). Recently, Sortilin has also been implicated in bindingto APP directly (Gustafsen, C et al., (2013) J Neurosc. 33:64-71) andalso to the APP processing enzyme BACE1 (Gustafsen, C et al., (2013) JNeurosc. 33:64-71; and Finan, G M et al., J Biol Chem 286: 12602-12616).Sortilin also binds to apolipoprotein E (APOE), to the A beta peptide(Carlo, A S et al., (2013) J Neurosc, 33: 358-370), and to PCSK9(Gustafsen et al, (2014) Cell Metab, 19: 310-318). Sortilin has alsobeen shown to bind to and regulate extracellular levels of PCSK9, whichdirects low-density lipoprotein receptor for degradation in lysosomes,resulting in increased levels of LDL cholesterol (Gustafsen C et al.,(2014). Cell Metab. 2014 Feb. 4; 19(2):310-8).

When present at intracellular vesicles such as endosomes, theamino-terminal extracellular domain of Sortilin is directed towards thelumen, where cargo of the vesicle is present. The carboxy-terminalintracellular/cytoplasmic domain of Sortilin, however, binds to a seriesof adaptor proteins, which regulate its trafficking from the surface andwithin intracellular compartments. These include AP2 (a clathrin adaptorto modulate endocytosis from the cell surface), and the RetromerComplex/AP1, which modulate movement from early endosomes to Golgi forrecycling; and interaction with GGA (Golgi-localizing, gamma-earcontaining, ADP-ribosylation factor binding) family proteins formovement from Golgi directly to early endosomes, usually for subsequentdegradation through lysosomes. Thus, Sortilin can bind to ligands at itsluminal domain, while engaging the cytoplasmic adaptors that determineits destination to determine intracellular fates, such as degradationfor Progranulin and other factors.

Through its various interactions with proteins, such as Progranulin,Sortilin and its multiple ligands have been shown to be involved invarious diseases, disorders, and conditions, such as frontotemporaldementia, amyotrophic lateral sclerosis, amyotrophic lateralsclerosis-frontotemporal dementia phenotypes, Alzheimer's disease,Parkinson's disease, depression, neuropsyciatric disorders, vasculardementia, seizures, retinal dystrophy, age related macular degeneration,glaucoma, traumatic brain injury, aging, seizures, wound healing,stroke, arthritis, and atherosclerotic vascular diseases.

Accordingly, there is a need for therapeutic antibodies thatspecifically bind Sortilin proteins and block the binding of Sortilin toits ligands, such as Progranulin, or otherwise modulate the effectiveconcentration of the ligands, in order to treat one or more diseases,disorders, and conditions associated with Sortilin activity.

All references cited herein, including patents, patent applications andpublications, are hereby incorporated by reference in their entirety.

SUMMARY

The present disclosure is generally directed to compositions thatinclude antibodies, e.g., monoclonal, chimeric, humanized antibodies,antibody fragments, etc., that specifically bind human Sortilin, and tomethods of using such compositions.

In some aspects, the present disclosure provides an antibody that bindsto a Sortilin protein, wherein the antibody comprises a heavy chainvariable region and a light chain variable region, wherein the heavychain variable region comprises an HVR-H1 comprising SEQ ID NO: 1, anHVR-H2 comprising SEQ ID NO: 2 or SEQ ID NO:3, and an HVR-H3 comprisingSEQ ID NO: 5 or SEQ ID NO: 6; and the light chain variable regioncomprises an HVR-L1 comprising any of SEQ ID NOs: 8-27, an HVR-L2comprising SEQ ID NO: 29 or SEQ ID NO: 30, and an HVR-L3 comprising SEQID NO: 32.

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the HVR-H1 comprisingthe amino acid sequence YSISSGYYWG (SEQ ID NO: 1), the HVR-H2 comprisingthe amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), the HVR-H3comprising the amino acid sequence ARQGSIQQGYYGMDV (SEQ ID NO: 5); and alight chain variable region comprising the HVR-L1 comprising the aminoacid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), the HVR-L2 comprising theamino acid sequence LGSNRAS (SEQ ID NO: 29), and the HVR-L3 comprisingthe amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the HVR-H1 comprisingthe amino acid sequence YSISSGYYWG (SEQ ID NO: 1), the HVR-H2 comprisingthe amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), the HVR-H3comprising the amino acid sequence ARQGSIQQGYYGMDV (SEQ ID NO: 5); and alight chain variable region comprising the HVR-L1 comprising the aminoacid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), the HVR-L2 comprising theamino acid sequence LGSNRVS (SEQ ID NO: 30), and the HVR-L3 comprisingthe amino acid sequence MQQQETPLT (SEQ ID NO: 33).

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the HVR-H1 comprisingthe amino acid sequence YSISSGYYWG (SEQ ID NO: 1), the HVR-H2 comprisingthe amino acid sequence TIYHSGSTYYNPSLES (SEQ ID NO: 3), the HVR-H3comprising the amino acid sequence ARQGSIQQGYYGMDV (SEQ ID NO: 5); and alight chain variable region comprising the HVR-L1 comprising the aminoacid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), the HVR-L2 comprising theamino acid sequence LGSNRAS (SEQ ID NO: 29), and the HVR-L3 comprisingthe amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some aspects, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the HVR-H1 comprisingthe amino acid sequence YSISSGYYWG (SEQ ID NO: 1), the HVR-H2 comprisingthe amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), the HVR-H3comprising the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and alight chain variable region comprising the HVR-L1 comprising the aminoacid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), the HVR-L2 comprising theamino acid sequence LGSNRAS (SEQ ID NO: 29), and the HVR-L3 comprisingthe amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some aspects, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the HVR-H1 comprisingthe amino acid sequence YSISSGYYWG (SEQ ID NO: 1), the HVR-H2 comprisingthe amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), the HVR-H3comprising the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and alight chain variable region comprising the HVR-L1 comprising the aminoacid sequence RSSQSLLRSTGYNYLD (SEQ ID NO: 9), the HVR-L2 comprising theamino acid sequence LGSNRAS (SEQ ID NO: 29), and the HVR-L3 comprisingthe amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some aspects, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the HVR-H1 comprisingthe amino acid sequence YSISSGYYWG (SEQ ID NO: 1), the HVR-H2 comprisingthe amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), the HVR-H3comprising the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and alight chain variable region comprising the HVR-L1 comprising the aminoacid sequence RSSQSLLRSTGYNYLD (SEQ ID NO: 10), the HVR-L2 comprisingthe amino acid sequence LGSNRAS (SEQ ID NO: 29), and the HVR-L3comprising the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some aspects, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the HVR-H1 comprisingthe amino acid sequence YSISSGYYWG (SEQ ID NO: 1), the HVR-H2 comprisingthe amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), the HVR-H3comprising the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and alight chain variable region comprising the HVR-L1 comprising the aminoacid sequence RSSQSLLRSTGYNYLD (SEQ ID NO: 21), the HVR-L2 comprisingthe amino acid sequence LGSNRAS (SEQ ID NO: 29), and the HVR-L3comprising the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the HVR-H1 comprisingthe amino acid sequence YSISSGYYWG (SEQ ID NO: 1), the HVR-H2 comprisingthe amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), the HVR-H3comprising the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and alight chain variable region comprising the HVR-L1 comprising the aminoacid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), the HVR-L2 comprising theamino acid sequence LGSNRAS (SEQ ID NO: 29), and the HVR-L3 comprisingthe amino acid sequence MQQQETPLT (SEQ ID NO: 33).

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the HVR-H1 comprisingthe amino acid sequence YSISSGYYWG (SEQ ID NO: 1), the HVR-H2 comprisingthe amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), the HVR-H3comprising the amino acid sequence ARQGSIQQGYYGMDV (SEQ ID NO: 5); and alight chain variable region comprising the HVR-L1 comprising the aminoacid sequence RSSQSLLHSNGYNYLD (SEQ ID NO: 26), the HVR-L2 comprisingthe amino acid sequence LGSNRAS (SEQ ID NO: 29), and the HVR-L3comprising the amino acid sequence MQQQETPLT (SEQ ID NO: 33).

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the HVR-H1 comprisingthe amino acid sequence YSISSGYYWG (SEQ ID NO: 1), the HVR-H2 comprisingthe amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), the HVR-H3comprising the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and alight chain variable region comprising the HVR-L1 comprising the aminoacid sequence RSSQGLLRSNGYNYLD (SEQ ID NO: 27), the HVR-L2 comprisingthe amino acid sequence LGSNRAS (SEQ ID NO: 29), and the HVR-L3comprising the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some aspects, the present disclosure provides an antibody that bindsto a Sortilin protein, wherein the antibody comprises a heavy chainvariable region and a light chain variable region, wherein the heavychain variable region comprises an HVR-H1 comprising the amino acidsequence YSISSGYYWG (SEQ ID NO: 1), an HVR-H2 comprising the amino acidsequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), and an HVR-H3 comprising theamino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and the light chainvariable region comprises an HVR-L1 comprising the amino acid sequenceRSSQSLLRSNGYNYLD (SEQ ID NO: 8), an HVR-L2 comprising the amino acidsequence LGSNRAS (SEQ ID NO: 29), and an HVR-L3 comprising the aminoacid sequence MQQQEAPLT (SEQ ID NO: 32).

In some aspects, the present disclosure provides an antibody that bindsto a Sortilin protein, wherein the antibody comprises a heavy chainvariable region and a light chain variable region, wherein the antibodycomprises a heavy chain variable region with an HVR-H1 comprising theamino acid sequence YSISSGYYWG (SEQ ID NO: 1), an HVR-H2 comprising theamino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), and an HVR-H3comprising the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); andthe light chain variable region comprises an HVR-L1 comprising the aminoacid sequence RSSQSLLRSTGYNYLD (SEQ ID NO: 9), an HVR-L2 comprising theamino acid sequence LGSNRAS (SEQ ID NO: 29), and an HVR-L3 comprisingthe amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some embodiments of any of the above aspects, the anti-Sortilinantibody: (a) decreases cell surface levels of Sortilin more than thelevel of decrease caused by an anti-Sortilin antibody comprising a heavychain variable region comprising the sequence of SEQ ID NO: 56 and alight chain variable region comprising the sequence of SEQ ID NO: 79;(b) increases extracellular levels of Progranulin more than the level ofincrease caused by an anti-Sortilin antibody comprising a heavy chainvariable region comprising the sequence of SEQ ID NO: 56 and a lightchain variable region comprising the sequence of SEQ ID NO: 79; (c)inhibits the interaction between Sortilin and Progranulin more than thelevel of inhibition caused by an anti-Sortilin antibody comprising aheavy chain variable region comprising the sequence of SEQ ID NO: 56 anda light chain variable region comprising the sequence of SEQ ID NO: 79;or (d) any combination of (a)-(c). In some embodiments of any of theabove aspects, the anti-Sortilin antibody: (a) decreases cell surfacelevels of Sortilin more than the level of decrease caused by ananti-Sortilin antibody selected from the group consisting of S-60-1,S-60-2, S-60-3, S-60-4, S-60-7, or S-60-8; (b) increases extracellularlevels of Progranulin more than the level of increase caused by ananti-Sortilin antibody selected from the group consisting of S-60-1,S-60-2, S-60-3, S-60-4, S-60-7, or S-60-8; (c) inhibits the interactionbetween Sortilin and Progranulin more than the level of inhibitioncaused by an anti-Sortilin antibody selected from the group consistingof S-60-1, S-60-2, S-60-3, S-60-4, S-60-7, or S-60-8; or (d) anycombination of (a)-(c).

Certain aspects of the present disclosure provide an anti-Sortilinantibody with a heavy chain variable region comprising an amino acidsequence selected from the group consisting of SEQ ID NOs: 54-56; and/ora light chain variable region comprising an amino acid sequence selectedfrom the group consisting of SEQ ID NOs: 57-58, 60-78, or 80.

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the amino acidsequence of SEQ ID NO: 54, and a light chain variable region comprisingthe amino acid sequence of SEQ ID NO: 57.

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the amino acidsequence of SEQ ID NO: 54, and a light chain variable region comprisingthe amino acid sequence of SEQ ID NO: 58.

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the amino acidsequence of SEQ ID NO: 54, and a light chain variable region comprisingthe amino acid sequence of SEQ ID NO: 59.

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the amino acidsequence of SEQ ID NO: 55, and a light chain variable region comprisingthe amino acid sequence of SEQ ID NO: 57.

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the amino acidsequence of SEQ ID NO: 55, and a light chain variable region comprisingthe amino acid sequence of SEQ ID NO: 58.

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the amino acidsequence of SEQ ID NO: 56, and a light chain variable region comprisingthe amino acid sequence of SEQ ID NO: 57.

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the amino acidsequence of SEQ ID NO: 56, and a light chain variable region comprisingthe amino acid sequence of SEQ ID NO: 77.

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the amino acidsequence of SEQ ID NO: 56, and a light chain variable region comprisingthe amino acid sequence of SEQ ID NO: 78.

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the amino acidsequence of SEQ ID NO: 54, and a light chain variable region comprisingthe amino acid sequence of SEQ ID NO: 79.

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the amino acidsequence of SEQ ID NO: 56, and a light chain variable region comprisingthe amino acid sequence of SEQ ID NO: 80.

In some aspects, the present disclosure provides an antibody that bindsto a Sortilin protein, wherein the antibody comprises a heavy chainvariable region comprising the amino acid sequence of SEQ ID NO: 56 anda light chain variable region comprising the amino acid sequence of SEQID NO: 57.

In certain aspects, the present disclosure provides an antibody thatbinds to a Sortilin protein, wherein the antibody comprises a heavychain variable region comprising the amino acid sequence of SEQ ID NO:56 and a light chain variable region comprising the amino acid sequenceof SEQ ID NO: 60.

In certain aspects, the present disclosure provides an antibody thatbinds to a Sortilin protein, wherein the antibody comprises a heavychain variable region comprising the amino acid sequence of SEQ ID NO:56 and a light chain variable region comprising the amino acid sequenceof SEQ ID NO: 61.

In certain aspects, the present disclosure provides an antibody thatbinds to a Sortilin protein, wherein the antibody comprises a heavychain variable region comprising the amino acid sequence of SEQ ID NO:56 and a light chain variable region comprising the amino acid sequenceof SEQ ID NO: 72.

In some embodiments that may be combined with any of the previousaspects, an anti-Sortilin antibody of the present disclosure is of theIgG class, the IgM class, or the IgA class. In certain specificembodiments, the anti-Sortilin antibody is of the IgG class and has anIgG1, IgG2, IgG3, or IgG4 isotype. In some embodiments of the presentdisclosure, provided is an anti-Sortilin antibody of the IgG1, IgG2,IgG3, or IgG4 isotype, wherein (a) the antibody is an IgG1 or IgG2isotype and the Fc region comprises an amino acid substitution atposition P331S, wherein the numbering of the residue position isaccording to EU numbering; (b) the antibody is an IgG1 isotype and theFc region comprises amino acid substitutions at positions L234A, L235A,and P331S, wherein the numbering of the residue position is according toEU numbering; (c) the antibody is an IgG1, IgG2, or IgG4 isotype and theFc region comprises an amino acid substitution at position N297A,wherein the numbering of the residue position is according to EUnumbering; or (d) the Fc region comprises an amino acid substitution atpositions S267E and L328F, wherein the numbering of the residue positionis according to EU numbering. In some embodiments, the antibody is anIgG1 isotype and the Fc region comprises amino acid substitutions atpositions L234A, L235A, and P331S, wherein the numbering of the residueposition is according to EU numbering.

In some aspects, the present disclosure provides an antibody that bindsto a Sortilin protein, wherein the antibody comprises a heavy chaincomprising the amino acid sequence of SEQ ID NO: 137 and a light chaincomprising the amino acid sequence of SEQ ID NO: 142.

In some aspects, the present disclosure provides an antibody that bindsto a Sortilin protein, wherein the antibody comprises a heavy chaincomprising the amino acid sequence of SEQ ID NO: 138 and a light chaincomprising the amino acid sequence of SEQ ID NO: 142.

In certain embodiments of the present disclosure, the Sortilin proteinis a human protein. In some embodiments, the Sortilin protein is awild-type protein. In certain embodiments, the Sortilin protein is anaturally occurring variant. In some embodiments, an anti-Sortilinantibody of the present disclosure binds specifically to a humanSortilin protein. In some embodiments that may be combined with any ofthe previous embodiments, the antibody is a human antibody, a bispecificantibody, a monoclonal antibody, a multivalent antibody, a conjugatedantibody, or a chimeric antibody. In certain embodiments, ananti-Sortilin antibody of the present disclosure is a monoclonalantibody.

In some embodiments, an anti-Sortilin antibody of the present disclosureis a bispecific antibody recognizing a first antigen and a secondantigen. In certain embodiments, bispecific antibodies of the presentdisclosure recognize both Sortilin and an antigen facilitating transportacross the blood-brain-barrier. In some specific embodiments, the firstantigen is Sortilin and the second antigen is Sortilin, transferrinreceptor (TR), insulin receptor (HIR), insulin-like growth factorreceptor (IGFR), low-density lipoprotein receptor related proteins 1 and2 (LPR-1 and 2), diphtheria toxin receptor, CRM197, a llama singledomain antibody, TMEM 30(A), a protein transduction domain, TAT, Syn-B,penetratin, a poly-arginine peptide, an angiopep peptide, basigin,Glut1, CD98hc, or ANG1005.

In certain embodiments of the present disclosure, the anti-Sortilinantibody is an antibody fragment that binds to a human Sortilin protein.In some embodiments, the anti-Sortilin antibody is an antibody fragmentthat binds to one or more human proteins selected from the groupconsisting of human Sortilin, a naturally occurring variant of humanSortilin, and a disease variant of human Sortilin. In certainembodiments that may be combined with any of the preceding embodiments,the anti-Sortilin antibody fragment is an Fab, Fab′, Fab′-SH, F(ab′)2,Fv, or scFv fragment.

In certain embodiments, the present disclosure provides an anti-Sortilinantibody, wherein (a) the anti-Sortilin antibody has increasesextracellular levels of Progranulin, decreases cellular levels ofSortilin, inhibits the interaction between Sortilin and Progranulin, orany combination thereof; (b) the anti-Sortilin antibody decreases cellsurface levels of Sortilin, increases extracellular levels ofProgranulin, inhibits interaction between Sortilin and Progranulin, orany combination thereof; (c) the anti-Sortilin antibody decreases cellsurface levels of Sortilin, decreases intracellular levels of Sortilin,decreases total levels of Sortilin, or any combination thereof; (d) theanti-Sortilin antibody induces Sortilin degradation, Sortilin cleavage,Sortilin internalization, Sortilin down regulation, or any combinationthereof; (e) the anti-Sortilin antibody decreases cellular levels ofSortilin and inhibits the interaction between Sortilin and Progranulin;(f) the anti-Sortilin antibody decreases cellular levels of Sortilin andincreases cellular levels of Progranulin; and/or (g) the anti-Sortilinantibody increases the effective concentration of Progranulin.

In certain embodiments, the present disclosure provides an anti-Sortilinantibody, wherein the anti-Sortilin antibody decreases cell surfacelevels of Sortilin, increases extracellular levels of Progranulin,inhibits interaction between Sortilin and Progranulin, or anycombination thereof. In some embodiments, the anti-Sortilin antibody hasa dissociation constant (K_(D)) for human Sortilin that is up to2.5-fold lower than an anti-Sortilin antibody comprising a heavy chainvariable region comprising the sequence of SEQ ID NO: 56 and a lightchain variable region comprising the sequence of SEQ ID NO: 79, whereinthe K_(D) is determined by FACS (see, e.g., Example 1). In someembodiments, the antibody has a dissociation constant (K_(D)) for humanSortilin that is greater than 1- and up to about 2.1-fold lower than ananti-Sortilin antibody comprising a heavy chain variable regioncomprising the sequence of SEQ ID NO: 56 and a light chain variableregion comprising the sequence of SEQ ID NO: 79, wherein the K_(D) isdetermined by FACS. In some embodiments, the anti-Sortilin antibody hasa dissociation constant (K_(D)) for human Sortilin that ranges fromabout 1.10E-8 M to about 4.68E-10 M wherein the K_(D) is determined byFACS (see, e.g., Example 1), or about 270 to about 2910 pM wherein theK_(D) is determined by Bio-layer interferometry (see, e.g., Example 4).In some embodiments, the antibody has a dissociation constant (K_(D))for human Sortilin that ranges from about 5.0E-10 M to about 1.0E-9 Mwherein the K_(D) is determined by FACS, or about 250-500 pM wherein theK_(D) is determined by Bio-layer interferometry.

In some embodiments that may be combined with any of the previousembodiments, an anti-Sortilin antibody of the present disclosure (a)reduces cell surface levels of Sortilin with a half maximal effectiveconcentration (EC₅₀) that is less than 150 pM, as measured by flowcytometry; (b) reduces cell surface levels of Sortilin by more thanabout 50% at 1.25 nM IgG, by more than about 80% at 0.63 nM IgG, or bymore than about 69% at 150 nM IgG relative to control, as measured byflow cytometry; increases Progranulin secretion by more than about 1.13fold over control at 0.63 nM IgG, or by more than about 1.22 fold overcontrol at 50 nM IgG, as measured by standard ELISA; blocks binding ofProgranulin to Sortilin with a half maximal effective concentration(EC₅₀) that is less than 0.325 nM, as measured by flow cytometry; (e)blocks binding of Progranulin to Sortilin by more than about 88% at 50nM IgG, or by more than about 27.5% at 150 nM IgG relative to control,as measured by flow cytometry; or (f) any combination thereof.

In some embodiments, an anti-Sortilin antibody of the present disclosure(a) reduces cell surface levels of Sortilin with a half maximaleffective concentration (EC₅₀) that is less than 681 pM, as measured byflow cytometry; (b) reduces cell surface levels of Sortilin by more thanabout 40% at 1.25 nM IgG, by more than about 29% at 0.6 nM IgG, or bymore than about 62% at 150 nM IgG relative to control, as measured byflow cytometry; (c) increases Progranulin secretion by more than about1.11 fold over control at 0.63 nM IgG, or by more than about 1.75 foldover control at 50 nM IgG, as measured by standard ELISA; (d) blocksbinding of Progranulin to Sortilin with a half maximal effectiveconcentration (EC₅₀) that is less than 0.751 nM, as measured by flowcytometry; (e) blocks binding of Progranulin to Sortilin by more thanabout 90% at 50 nM IgG, or by more than about 95% at 150 nM IgG relativeto control, as measured by flow cytometry; or (f) any combinationthereof.

In some embodiments, an anti-Sortilin antibody of the present disclosurecompetes with an antibody comprising the heavy chain variable domain andthe light chain variable domain of an antibody selected from the groupconsisting of S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33(wt)], S-60-15.1 [N33T], S-60-16, S-60-18, S-60-19, S-60-24, and anycombination thereof for binding to Sortilin. In some embodiments thatmay be combined with any of the previous embodiments, an anti-Sortilinantibody of the present disclosure binds essentially the same Sortilinepitope as an antibody comprising the heavy chain variable domain andthe light chain variable domain of an antibody selected from the groupconsisting of S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33(wt)], S-60-15.1 [N33T], S-60-16, S-60-18, S-60-19, and S-60-24.

In another aspect, the present disclosure relates to an antibodyproduced by any of the methods described herein. In some aspects, thepresent disclosure provides an isolated nucleic acid comprising anucleic acid sequence encoding an anti-Sortilin antibody of any one ofthe preceding aspects and embodiments. Accordingly, some aspects providea vector containing a nucleic acid encoding an anti-Sortilin antibody,and some aspects provide an isolated host cell containing such a vector.In certain aspects, the present disclosure provides a method ofproducing an anti-Sortilin antibody that binds to Sortilin, comprisingculturing such an isolated host cell so that the anti-Sortilin antibodyis produced. In certain embodiments, the method further comprisesrecovering the antibody produced by the cell. In some aspects, provideherein is an isolated antibody that binds to Sortilin and is produced bya method of the present disclosure.

In another aspect, the present disclosure relates to a pharmaceuticalcomposition comprising any of the anti-Sortilin antibodies describedherein and a pharmaceutically acceptable carrier. In some embodiments,the present disclosure relates to the use of any of the antibodies orcompositions described herein for the preparation or manufacture of amedicament. In some embodiments, the present disclosure relates to theuse of any of the antibodies or compositions described herein for thepreparation or manufacture of a medicament for the treatment offrontotemporal dementia, progressive supranuclear palsy, Alzheimer'sdisease, vascular dementia, seizures, retinal dystrophy, amyotrophiclateral sclerosis, traumatic brain injury, a spinal cord injury,neuropathic pain, dementia, stroke, Parkinson's disease, acutedisseminated encephalomyelitis, retinal degeneration, age relatedmacular degeneration, glaucoma, multiple sclerosis, septic shock,bacterial infection, arthritis, or osteoarthritis. Some embodimentsprovide an antibody or composition described herein for use in a methodof treatment. Some embodiments provide an antibody or compositiondescribed herein for use in a method of treatment of frontotemporaldementia, progressive supranuclear palsy, Alzheimer's disease, vasculardementia, seizures, retinal dystrophy, amyotrophic lateral sclerosis,traumatic brain injury, a spinal cord injury, neuropathic pain,dementia, stroke, Parkinson's disease, acute disseminatedencephalomyelitis, retinal degeneration, age related maculardegeneration, glaucoma, multiple sclerosis, septic shock, bacterialinfection, arthritis, or osteoarthritis.

In certain aspects, provided herein is a method of preventing, reducingrisk for, or treating an individual having a disease, disorder, orinjury, comprising administering to an individual in need thereof atherapeutically effective amount of an anti-Sortilin antibody of thepresent disclosure. In some embodiments, the disease, disorder or injuryis selected from the group consisting of frontotemporal dementia,progressive supranuclear palsy, Alzheimer's disease, vascular dementia,seizures, retinal dystrophy, amyotrophic lateral sclerosis, traumaticbrain injury, a spinal cord injury, neuropathic pain, dementia, stroke,Parkinson's disease, acute disseminated encephalomyelitis, retinaldegeneration, age related macular degeneration, glaucoma, multiplesclerosis, septic shock, bacterial infection, arthritis, andosteoarthritis. In some embodiments, the disease, disorder, or injury isselected from frontotemporal dementia, progressive supranuclear palsy,Alzheimer's disease, and amyotrophic lateral sclerosis.

In certain aspects, provided herein is a method of inhibiting one ormore of neuroinflammation, axonopathy characterized by short axonaloutgrowth and aberrant branching, microglial activation, andinflammatory response, comprising administering to the individual atherapeutically effective amount of an anti-Sortilin antibody of thepresent disclosure.

In certain aspects, provided herein is a method of promoting one or moreof wound healing, autophagy, and clearance of aggregate proteins,comprising administering to the individual a therapeutically effectiveamount of an anti-Sortilin antibody of the present disclosure.

In certain aspects, provided herein is a method of preventing, reducingrisk, or treating an individual having arthritis, comprisingadministering to the individual a therapeutically effective amount of ananti-Sortilin antibody of the present disclosure.

In certain aspects, provided herein is a method of decreasing expressionof one or more pro-inflammatory mediators, comprising administering tothe individual a therapeutically effective amount of an anti-Sortilinantibody of the present disclosure. In some embodiments, the one or morepro-inflammatory mediators are selected from the group consisting ofIL-6, IL12p70, IL12p40, IL-1β, TNF-α, CXCL1, CCL2, CCL3, CCL4, and CCL5.

In some aspects, provided herein is a method of inhibiting theinteraction between Sortilin and Progranulin, the method comprisingexposing a cell expressing Sortilin to an anti-Sortilin antibody or apharmaceutical composition of the present disclosure. In someembodiments, the cell is in vitro. In some embodiments, the cell is invivo. In some embodiments, the method further comprises decreasing thelevel of Sortilin expressed on the cell surface. In some embodiments,extracellular levels of Progranulin are increased.

In some aspects, provided herein is a method of increasing levels ofProgranulin in an individual in need thereof, the method comprisingadministering to the individual a therapeutically effective amount ofthe anti-Sortilin antibody or a pharmaceutical composition of thepresent disclosure. In some embodiments, levels of Progranulin areincreased in plasma. In some embodiments, levels of Progranulin areincreased in cerebrospinal fluid. In some aspects, provided herein is amethod of decreasing levels of Sortilin in an individual in needthereof, the method comprising administering to the individual atherapeutically effective amount of an anti-Sortilin antibody or apharmaceutical composition of the present disclosure. In someembodiments, the levels of Sortilin are decreased in peripheral whiteblood cells. In some embodiments, the individual has one or moremutations in the gene encoding Progranulin. In some the individual isheterozygous for one or more loss-of-function mutations in the geneencoding Progranulin. In some embodiments, the individual has a c9orf72hexanucleotide repeat expansion. the individual has or is at risk forfrontotemporal dementia, Alzheimer's disease, or amyotrophic lateralsclerosis.

In some embodiments that may be combined with any of the previousaspects or embodiments, the present disclosure provides a method,wherein the anti-Sortilin antibody comprises two or more anti-Sortilinantibodies.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A-1C provide pharmacokinetic and pharmacodynamic studies ofnon-human primates administered single doses of anti-Sortilin antibodyS-60-15.1 [N33T] LALAPS (S-60-15.1 [N33T] with huIgG1 withL234A/L235A/P331S mutations). FIG. 1A provides the level of SORT1 inperipheral white blood cells as a percentage from baseline at theindicated times after treatment (hours) with the specified anti-Sortilinantibody doses. SORT1 expression decreased with all of the anti-Sortilinantibody doses tested. Higher antibody doses (60 mg/kg, 200 mg/kg)resulted in both an earlier and more prolonged decrease of SORT1 levelscompared to lower anti-Sortilin antibody doses (5 mg/kg, 20 mg/kg). FIG.1B provides the levels of PGRN in the plasma as a percentage frombaseline at the indicated times after treatment (hours) with thespecified anti-Sortilin antibody doses. The levels of PGRN increased ina time- and dose-dependent manner. In particular, plasma PGRN levelsincreased 3- to 4-fold at C_(max), compared to baseline levels, for allanti-Sortilin antibody doses tested and remained elevated for longerperiods of time at the higher antibody doses. FIG. 1C provides thelevels of PGRN in CSF as a percentage from baseline at the indicatedtimes after treatment (hours) with the specified anti-Sortilin antibodydoses. CSF PGRN levels increased 2- to 3-fold above baseline in animalsadministered either 20 mg/kg, 60 mg/kg, or 200 mg/kg. As observed withplasma PGRN levels (FIG. 1B), CSF PGRN levels remained elevated overtime in the higher antibody dose groups. For FIGS. 1A-1C, n=3 animalsper dose.

FIGS. 2A-2C provide pharmacokinetic and pharmacodynamic studies ofnon-human primates administered repeat doses of anti-Sortilin antibodyS-60-15.1 [N33T] LALAPS. Animals (2 males and 2 females) wereadministered anti-Sortilin antibody S-60-15.1 [N33T] LALAPS at a dose of60 mg/kg once per week for four weeks. The days on which dosing occurredare represented by the vertical dashed lines. FIG. 2A provides the mean(+/− standard deviation) of the concentration of SORT1 in peripheralwhite blood cells (WBCs) as a percentage of baseline at the indicatedtimes (days). SORT1 levels in peripheral white blood cells remaineddecreased throughout the duration of the study.

FIG. 2B provides the mean (+/− standard deviation) of the concentrationof PGRN in plasma as a percentage of baseline (normalized) at theindicated times (days). Plasma PGRN levels increased to 5- to 6-foldabove baseline at peak levels. A decrease in plasma PGRN was observedfollowing the fourth and final administration of anti-Sortilin antibody;however, the plasma PGRN levels remained elevated by 2-fold abovebaseline. FIG. 2C provides the mean (+/− standard deviation) of theconcentration of PGRN in CSF as a percentage of baseline (normalized) atthe indicated times (days). CSF PGRN levels were increased 3- to 4-foldabove baseline (FIG. 2C).

DETAILED DESCRIPTION Definitions

As used herein, the term “preventing” includes providing prophylaxiswith respect to occurrence or recurrence of a particular disease,disorder, or condition in an individual. An individual may bepredisposed to, susceptible to a particular disease, disorder, orcondition, or at risk of developing such a disease, disorder, orcondition, but has not yet been diagnosed with the disease, disorder, orcondition.

As used herein, an individual “at risk” of developing a particulardisease, disorder, or condition may or may not have detectable diseaseor symptoms of disease, and may or may not have displayed detectabledisease or symptoms of disease prior to the treatment methods describedherein. “At risk” denotes that an individual has one or more riskfactors, which are measurable parameters that correlate with developmentof a particular disease, disorder, or condition, as known in the art. Anindividual having one or more of these risk factors has a higherprobability of developing a particular disease, disorder, or conditionthan an individual without one or more of these risk factors.

As used herein, the term “treatment” refers to clinical interventiondesigned to alter the natural course of the individual being treatedduring the course of clinical pathology. Desirable effects of treatmentinclude decreasing the rate of progression, ameliorating or palliatingthe pathological state, and remission or improved prognosis of aparticular disease, disorder, or condition. An individual issuccessfully “treated”, for example, if one or more symptoms associatedwith a particular disease, disorder, or condition are mitigated oreliminated.

An “effective amount” refers to at least an amount effective, at dosagesand for periods of time necessary, to achieve the desired therapeutic orprophylactic result. An effective amount can be provided in one or moreadministrations. An effective amount herein may vary according tofactors such as the disease state, age, sex, and weight of theindividual, and the ability of the treatment to elicit a desiredresponse in the individual. An effective amount is also one in which anytoxic or detrimental effects of the treatment are outweighed by thetherapeutically beneficial effects. For prophylactic use, beneficial ordesired results include results such as eliminating or reducing therisk, lessening the severity, or delaying the onset of the disease,including biochemical, histological and/or behavioral symptoms of thedisease, its complications and intermediate pathological phenotypespresenting during development of the disease. For therapeutic use,beneficial or desired results include clinical results such asdecreasing one or more symptoms resulting from the disease, increasingthe quality of life of those suffering from the disease, decreasing thedose of other medications required to treat the disease, enhancingeffect of another medication such as via targeting, delaying theprogression of the disease, and/or prolonging survival. An effectiveamount of drug, compound, or pharmaceutical composition is an amountsufficient to accomplish prophylactic or therapeutic treatment eitherdirectly or indirectly. As is understood in the clinical context, aneffective amount of a drug, compound, or pharmaceutical composition mayor may not be achieved in conjunction with another drug, compound, orpharmaceutical composition. Thus, an “effective amount” may beconsidered in the context of administering one or more therapeuticagents, and a single agent may be considered to be given in an effectiveamount if, in conjunction with one or more other agents, a desirableresult may be or is achieved

As used herein, administration “in conjunction” with another compound orcomposition includes simultaneous administration and/or administrationat different times. Administration in conjunction also encompassesadministration as a co-formulation or administration as separatecompositions, including at different dosing frequencies or intervals,and using the same route of administration or different routes ofadministration.

An “individual” for purposes of treatment, prevention, or reduction ofrisk refers to any animal classified as a mammal, including humans,domestic and farm animals, and zoo, sport, or pet animals, such as dogs,horses, rabbits, cattle, pigs, hamsters, gerbils, mice, ferrets, rats,cats, and the like. Preferably, the individual is human.

The terms “Sortilin” or “Sortilin polypeptide” are used interchangeablyherein refer herein to any native Sortilin from any mammalian source,including primates (e.g., humans and cynos) and rodents (e.g., mice andrats), unless otherwise indicated. In some embodiments, the termencompasses both wild-type sequences and naturally occurring variantsequences, e.g., splice variants or allelic variants. In someembodiments, the term encompasses “full-length,” unprocessed Sortilin aswell as any form of Sortilin that results from processing in the cell.In some embodiments, the Sortilin is human Sortilin. In someembodiments, the amino acid sequence of an exemplary human Sortilin isSEQ ID NO: 81.

The terms “anti-Sortilin antibody,” an “antibody that binds toSortilin,” and “antibody that specifically binds Sortilin” refer to anantibody that is capable of binding Sortilin with sufficient affinitysuch that the antibody is useful as a diagnostic and/or therapeuticagent in targeting Sortilin. In one embodiment, the extent of binding ofan anti-Sortilin antibody to an unrelated, non-Sortilin polypeptide isless than about 10% of the binding of the antibody to Sortilin asmeasured, e.g., by a radioimmunoassay (RIA). In certain embodiments, anantibody that binds to Sortilin has a dissociation constant (K_(D)) of<1 μM, <100 nM, <10 nM, <1 nM, <0.1 nM, <0.01 nM, or <0.001 nM (e.g.,10-8 M or less, e.g. from 10-8 M to 10-13 M, e.g., from 10-9 M to 10-13M). In certain embodiments, an anti-Sortilin antibody binds to anepitope of Sortilin that is conserved among Sortilin from differentspecies.

The term “immunoglobulin” (Ig) is used interchangeably with “antibody”herein. The term “antibody” herein is used in the broadest sense andspecially covers monoclonal antibodies, polyclonal antibodies,multispecific antibodies (e.g., bispecific antibodies) including thoseformed from at least two intact antibodies, and antibody fragments solong as they exhibit the desired biological activity.

“Native antibodies” are usually heterotetrameric glycoproteins of about150,000 Daltons, composed of two identical Light (“L”) chains and twoidentical heavy (“H”) chains. Each light chain is linked to a heavychain by one covalent disulfide bond, while the number of disulfidelinkages varies among the heavy chains of different immunoglobulinisotypes. Each heavy and light chain also has regularly spacedintra-chain disulfide bridges. Each heavy chain has at one end avariable domain (V_(H)) followed by a number of constant domains. Eachlight chain has a variable domain at one end (V_(L)) and a constantdomain at its other end; the constant domain of the light chain isaligned with the first constant domain of the heavy chain, and the lightchain variable domain is aligned with the variable domain of the heavychain. Particular amino acid residues are believed to form an interfacebetween the light chain and heavy chain variable domains.

For the structure and properties of the different classes of antibodies,see, e.g., Basic and Clinical Immunology, 8th Ed., Daniel P. Stites,Abba I. Terr and Tristram G. Parslow (eds.), Appleton & Lange, Norwalk,Conn., 1994, page 71 and Chapter 6.

The light chain from any vertebrate species can be assigned to one oftwo clearly distinct types, called kappa (“κ”) and lambda (“λ”), basedon the amino acid sequences of their constant domains. Depending on theamino acid sequence of the constant domain of their heavy chains (CH),immunoglobulins can be assigned to different classes or isotypes. Thereare five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, havingheavy chains designated alpha (“α”), delta (“δ”), epsilon (“ε”), gamma(“γ”), and mu (“μ”), respectively. The γ and α classes are furtherdivided into subclasses (isotypes) on the basis of relatively minordifferences in the CH sequence and function, e.g., humans express thefollowing subclasses: IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. Thesubunit structures and three-dimensional configurations of differentclasses of immunoglobulins are well known and described generally in,for example, Abbas et al., Cellular and Molecular Immunology, 4^(th) ed.(W.B. Saunders Co., 2000).

The “variable region” or “variable domain” of an antibody, such as ananti-Sortilin antibody of the present disclosure, refers to theamino-terminal domains of the heavy or light chain of the antibody. Thevariable domains of the heavy chain and light chain may be referred toas “V_(H)” and “V_(L)”, respectively. These domains are generally themost variable parts of the antibody (relative to other antibodies of thesame class) and contain the antigen binding sites.

The term “variable” refers to the fact that certain segments of thevariable domains differ extensively in sequence among antibodies, suchas anti-Sortilin antibodies of the present disclosure. The variabledomain mediates antigen binding and defines the specificity of aparticular antibody for its particular antigen. However, the variabilityis not evenly distributed across the entire span of the variabledomains. Instead, it is concentrated in three segments calledhypervariable regions (HVRs) both in the light-chain and the heavy chainvariable domains. The more highly conserved portions of variable domainsare called the framework regions (FR). The variable domains of nativeheavy and light chains each comprise four FR regions, largely adopting abeta-sheet configuration, connected by three HVRs, which form loopsconnecting, and in some cases forming part of, the beta-sheet structure.The HVRs in each chain are held together in close proximity by the FRregions and, with the HVRs from the other chain, contribute to theformation of the antigen-binding site of antibodies (see Kabat et al.,Sequences of Immunological Interest, Fifth Edition, National Instituteof Health, Bethesda, Md. (1991)). The constant domains are not involveddirectly in the binding of antibody to an antigen, but exhibit variouseffector functions, such as participation of the antibody inantibody-dependent-cellular toxicity.

An “isolated” antibody, such as an anti-Sortilin antibody of the presentdisclosure, is one that has been identified, separated and/or recoveredfrom a component of its production environment (e.g., naturally orrecombinantly). Preferably, the isolated polypeptide is free ofassociation with all other contaminant components from its productionenvironment. Contaminant components from its production environment,such as those resulting from recombinant transfected cells, arematerials that would typically interfere with research, diagnostic ortherapeutic uses for the antibody, and may include enzymes, hormones,and other proteinaceous or non-proteinaceous solutes. In preferredembodiments, the polypeptide will be purified: (1) to greater than 95%by weight of antibody as determined by, for example, the Lowry method,and in some embodiments, to greater than 99% by weight; (2) to a degreesufficient to obtain at least 15 residues of N-terminal or internalamino acid sequence by use of a spinning cup sequenator, or (3) tohomogeneity by SDS-PAGE under non-reducing or reducing conditions usingCoomassie blue or, preferably, silver stain. Isolated antibody includesthe antibody in situ within recombinant T cells since at least onecomponent of the antibody's natural environment will not be present.Ordinarily, however, an isolated polypeptide or antibody will beprepared by at least one purification step.

The term “monoclonal antibody” as used herein refers to an antibody,such as a monoclonal anti-Sortilin antibody of the present disclosure,obtained from a population of substantially homogeneous antibodies,i.e., the individual antibodies comprising the population are identicalexcept for possible naturally occurring mutations and/orpost-translation modifications (e.g., isomerizations, amidations, etc.)that may be present in minor amounts. Monoclonal antibodies are highlyspecific, being directed against a single antigenic site. In contrast topolyclonal antibody preparations which typically include differentantibodies directed against different determinants (epitopes), eachmonoclonal antibody is directed against a single determinant on theantigen. In addition to their specificity, the monoclonal antibodies areadvantageous in that they are synthesized by the hybridoma culture,uncontaminated by other immunoglobulins. The modifier “monoclonal”indicates the character of the antibody as being obtained from asubstantially homogeneous population of antibodies, and is not to beconstrued as requiring production of the antibody by any particularmethod. For example, the monoclonal antibodies to be used in accordancewith the present invention may be made by a variety of techniques,including, but not limited to one or more of the following methods,immunization methods of animals including, but not limited to rats,mice, rabbits, guinea pigs, hamsters and/or chickens with one or more ofDNA(s), virus-like particles, polypetide(s), and/or cell(s), thehybridoma methods, B-cell cloning methods, recombinant DNA methods, andtechnologies for producing human or human-like antibodies in animalsthat have parts or all of the human immunoglobulin loci or genesencoding human immunoglobulin sequences.

The terms “full-length antibody,” “intact antibody” or “whole antibody”are used interchangeably to refer to an antibody, such as ananti-Sortilin antibody of the present disclosure, in its substantiallyintact form, as opposed to an antibody fragment. Specifically, wholeantibodies include those with heavy and light chains including an Fcregion. The constant domains may be native sequence constant domains(e.g., human native sequence constant domains) or amino acid sequencevariants thereof. In some cases, the intact antibody may have one ormore effector functions.

An “antibody fragment” comprises a portion of an intact antibody,preferably the antigen binding and/or the variable region of the intactantibody. Examples of antibody fragments include Fab, Fab′, F(ab′)₂ andFv fragments; diabodies; linear antibodies (see U.S. Pat. No. 5,641,870,Example 2; Zapata et al., Protein Eng. 8(10):1057-1062 (1995));single-chain antibody molecules and multispecific antibodies formed fromantibody fragments.

Papain digestion of antibodies, such as anti-Sortilin antibodies of thepresent disclosure, produces two identical antigen-binding fragments,called “Fab” fragments, and a residual “Fc” fragment, a designationreflecting the ability to crystallize readily. The Fab fragment consistsof an entire L chain along with the variable region domain of the Hchain (V_(H)), and the first constant domain of one heavy chain(C_(H)1). Each Fab fragment is monovalent with respect to antigenbinding, i.e., it has a single antigen-binding site. Pepsin treatment ofan antibody yields a single large F(ab′)₂ fragment which roughlycorresponds to two disulfide linked Fab fragments having differentantigen-binding activity and is still capable of cross-linking antigen.Fab′ fragments differ from Fab fragments by having a few additionalresidues at the carboxy terminus of the C_(H)1 domain including one ormore cysteines from the antibody hinge region. Fab′-SH is thedesignation herein for Fab′ in which the cysteine residue(s) of theconstant domains bear a free thiol group. F(ab′)₂ antibody fragmentsoriginally were produced as pairs of Fab′ fragments which have hingecysteines between them. Other chemical couplings of antibody fragmentsare also known.

The Fc fragment comprises the carboxy-terminal portions of both H chainsheld together by disulfides. The effector functions of antibodies aredetermined by sequences in the Fc region, the region which is alsorecognized by Fc receptors (FcR) found on certain types of cells.

“Fv” is the minimum antibody fragment which contains a completeantigen-recognition and -binding site. This fragment consists of a dimerof one heavy- and one light-chain variable region domain in tight,non-covalent association. From the folding of these two domains emanatesix hypervariable loops (3 loops each from the H and L chain) thatcontribute the amino acid residues for antigen binding and conferantigen binding specificity to the antibody. However, even a singlevariable domain (or half of an Fv comprising only three HVRs specificfor an antigen) has the ability to recognize and bind antigen, althoughat a lower affinity than the entire binding site.

“Single-chain Fv” also abbreviated as “sFv” or “scFv” are antibodyfragments that comprise the VH and VL antibody domains connected into asingle polypeptide chain. Preferably, the sFv polypeptide furthercomprises a polypeptide linker between the V_(H) and V_(L) domains whichenables the sFv to form the desired structure for antigen binding. For areview of the sFv, see Pluckthun in The Pharmacology of MonoclonalAntibodies, vol. 113, Rosenburg and Moore eds., Springer-Verlag, NewYork, pp. 269-315 (1994).

“Functional fragments” of antibodies, such as anti-Sortilin antibodiesof the present disclosure, comprise a portion of an intact antibody,generally including the antigen binding or variable region of the intactantibody or the F region of an antibody which retains or has modifiedFcR binding capability. Examples of antibody fragments include linearantibody, single-chain antibody molecules and multispecific antibodiesformed from antibody fragments.

The term “diabodies” refers to small antibody fragments prepared byconstructing sFv fragments (see preceding paragraph) with short linkers(about 5-10) residues) between the V_(H) and V_(L) domains such thatinter-chain but not intra-chain pairing of the variable domains isachieved, thereby resulting in a bivalent fragment, i.e., a fragmenthaving two antigen-binding sites. Bispecific diabodies are heterodimersof two “crossover” sFv fragments in which the V_(H) and VL domains ofthe two antibodies are present on different polypeptide chains.

As used herein, a “chimeric antibody” refers to an antibody(immunoglobulin), such as a chimeric anti-Sortilin antibody of thepresent disclosure, in which a portion of the heavy and/or light chainis identical with or homologous to corresponding sequences in antibodiesderived from a particular species or belonging to a particular antibodyclass or subclass, while the remainder of the chain(s) is(are) identicalwith or homologous to corresponding sequences in antibodies derived fromanother species or belonging to another antibody class or subclass, aswell as fragments of such antibodies, so long as they exhibit thedesired biological activity. Chimeric antibodies of interest hereininclude PRIMATIZED® antibodies wherein the antigen-binding region of theantibody is derived from an antibody produced by, e.g., immunizingmacaque monkeys with an antigen of interest. As used herein, “humanizedantibody” is used a subset of “chimeric antibodies.”

“Humanized” forms of non-human (e.g., murine) antibodies, such ashumanized forms of anti-Sortilin antibodies of the present disclosure,are chimeric antibodies comprising amino acid residues from non-humanHVRs and amino acid residues from human FRs. In certain embodiments, ahumanized antibody will comprise substantially all of at least one, andtypically two, variable domains, in which all or substantially all ofthe HVRs (e.g., CDRs) correspond to those of a non-human antibody, andall or substantially all of the FRs correspond to those of a humanantibody. A humanized antibody optionally may comprise at least aportion of an antibody constant region derived from a human antibody. A“humanized form” of an antibody, e.g., a non-human antibody, refers toan antibody that has undergone humanization.

A “human antibody” is one that possesses an amino-acid sequencecorresponding to that of an antibody, such as an anti-Sortilin antibodyof the present disclosure, produced by a human and/or has been madeusing any of the techniques for making human antibodies as disclosedherein. This definition of a human antibody specifically excludes ahumanized antibody comprising non-human antigen-binding residues. Humanantibodies can be produced using various techniques known in the art,including phage-display libraries and yeast-based platform technologies.Human antibodies can be prepared by administering the antigen to atransgenic animal that has been modified to produce such antibodies inresponse to antigenic challenge, but whose endogenous loci have beendisabled, e.g., immunized xenomice as well as generated via a humanB-cell hybridoma technology.

The term “hypervariable region,” “HVR,” or “HV,” when used herein refersto the regions of an antibody-variable domain, such as that of ananti-Sortilin antibody of the present disclosure, that are hypervariablein sequence and/or form structurally defined loops. Generally,antibodies comprise six HVRs; three in the V_(H) (H1, H2, H3), and threein the V_(L) (L1, L2, L3). In native antibodies, H3 and L3 display themost diversity of the six HVRs, and H3 in particular is believed to playa unique role in conferring fine specificity to antibodies. Naturallyoccurring camelid antibodies consisting of a heavy chain only arefunctional and stable in the absence of light chain.

A number of HVR delineations are in use and are encompassed herein. Insome embodiments, the HVRs may be Kabat complementarity-determiningregions (CDRs) based on sequence variability and are the most commonlyused (Kabat et al., supra). In some embodiments, the HVRs may be ChothiaCDRs. Chothia refers instead to the location of the structural loops(Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). In someembodiments, the HVRs may be AbM HVRs. The AbM HVRs represent acompromise between the Kabat CDRs and Chothia structural loops, and areused by Oxford Molecular's AbM antibody-modeling software. In someembodiments, the HVRs may be “contact” HVRs. The “contact” HVRs arebased on an analysis of the available complex crystal structures. Theresidues from each of these HVRs are noted below.

Loop Kabat AbM Chothia Contact L1 L24-L34 L24-L34 L26-L32 L30-L36 L2L50-L56 L50-L56 L50-L52 L46-L55 L3 L89-L97 L89-L97 L91-L96 L89-L96 H1H31-H35B H26-H35B H26-H32 H30-H35B (Kabat numbering) H1 H31-H35 H26-H35H26-H32 H30-H35 (Chothia numbering) H2 H50-H65 H50-H58 H53-H55 H47-H58H3 H95-H102 H95-H102 H96-H101 H93-H101

HVRs may comprise “extended HVRs” as follows: 24-36 or 24-34 (L1), 46-56or 50-56 (L2), and 89-97 or 89-96 (L3) in the VL, and 26-35 (H1), 50-65or 49-65 (a preferred embodiment) (H2), and 93-102, 94-102, or 95-102(H3) in the VH. The variable-domain residues are numbered according toKabat et al., supra, for each of these extended-HVR definitions.

“Framework” or “FR” residues are those variable-domain residues otherthan the HVR residues as herein defined.

An “acceptor human framework” as used herein is a framework comprisingthe amino acid sequence of a V_(L) or V_(H) framework derived from ahuman immunoglobulin framework or a human consensus framework. Anacceptor human framework “derived from” a human immunoglobulin frameworkor a human consensus framework may comprise the same amino acid sequencethereof, or it may comprise pre-existing amino acid sequence changes. Insome embodiments, the number of pre-existing amino acid changes are 10or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 orless, 3 or less, or 2 or less. Where pre-existing amino acid changes arepresent in a VH, preferable those changes occur at only three, two, orone of positions 71H, 73H and 78H; for instance, the amino acid residuesat those positions may by 71A, 73T and/or 78A. In one embodiment, the VLacceptor human framework is identical in sequence to the V_(L) humanimmunoglobulin framework sequence or human consensus framework sequence.

A “human consensus framework” is a framework that represents the mostcommonly occurring amino acid residues in a selection of humanimmunoglobulin V_(L) or V_(H) framework sequences. Generally, theselection of human immunoglobulin V_(L) or V_(H) sequences is from asubgroup of variable domain sequences. Generally, the subgroup ofsequences is a subgroup as in Kabat et al., Sequences of Proteins ofImmunological Interest, 5th Ed. Public Health Service, NationalInstitutes of Health, Bethesda, Md. (1991). Examples include for theV_(L), the subgroup may be subgroup kappa I, kappa II, kappa III orkappa IV as in Kabat et al., supra. Additionally, for the V_(H), thesubgroup may be subgroup I, subgroup II, or subgroup III as in Kabat etal., supra.

An “amino-acid modification” at a specified position, e.g., of ananti-Sortilin antibody of the present disclosure, refers to thesubstitution or deletion of the specified residue, or the insertion ofat least one amino acid residue adjacent the specified residue.Insertion “adjacent” to a specified residue means insertion within oneto two residues thereof. The insertion may be N-terminal or C-terminalto the specified residue. The preferred amino acid modification hereinis a substitution.

An “affinity-matured” antibody, such as an anti-Sortilin antibody of thepresent disclosure, is one with one or more alterations in one or moreHVRs thereof that result in an improvement in the affinity of theantibody for antigen, compared to a parent antibody that does notpossess those alteration(s). In one embodiment, an affinity-maturedantibody has nanomolar or even picomolar affinities for the targetantigen. Affinity-matured antibodies are produced by procedures known inthe art. For example, Marks et al., Bio/Technology 10:779-783 (1992)describes affinity maturation by VH- and VL-domain shuffling. Randommutagenesis of HVR and/or framework residues is described by, forexample: Barbas et al. Proc Nat. Acad. Sci. USA 91:3809-3813 (1994);Schier et al. Gene 169:147-155 (1995); Yelton et al. J. Immunol.155:1994-2004 (1995); Jackson et al., J. Immunol. 154(7):3310-9 (1995);and Hawkins et al, J. Mol. Biol. 226:889-896 (1992).

As use herein, the term “specifically recognizes” or “specificallybinds” refers to measurable and reproducible interactions such asattraction or binding between a target and an antibody, such as ananti-Sortilin antibody of the present disclosure, that is determinativeof the presence of the target in the presence of a heterogeneouspopulation of molecules including biological molecules. For example, anantibody, such as an anti-Sortilin antibody of the present disclosure,that specifically or preferentially binds to a target or an epitope isan antibody that binds this target or epitope with greater affinity,avidity, more readily, and/or with greater duration than it binds toother targets or other epitopes of the target. It is also understood byreading this definition that, for example, an antibody (or a moiety)that specifically or preferentially binds to a first target may or maynot specifically or preferentially bind to a second target. As such,“specific binding” or “preferential binding” does not necessarilyrequire (although it can include) exclusive binding. An antibody thatspecifically binds to a target may have an association constant of atleast about 10³ M⁻¹ or 10⁴ M⁻¹, sometimes about 10⁵ M⁻¹ or 10⁶ M⁻¹, inother instances about 10⁶ M⁻¹ or 10⁷ M⁻¹, about 10⁸ M⁻¹ to 10⁹ M⁻¹, orabout 10¹⁰ M⁻¹ to 10¹¹ M⁻¹ or higher. A variety of immunoassay formatscan be used to select antibodies specifically immunoreactive with aparticular protein. For example, solid-phase ELISA immunoassays areroutinely used to select monoclonal antibodies specificallyimmunoreactive with a protein. See, e.g., Harlow and Lane (1988)Antibodies, A Laboratory Manual, Cold Spring Harbor Publications, NewYork, for a description of immunoassay formats and conditions that canbe used to determine specific immunoreactivity.

As used herein, an “interaction” between a Sortilin protein and a secondprotein encompasses, without limitation, protein-protein interaction, aphysical interaction, a chemical interaction, binding, covalent binding,and ionic binding. As used herein, an antibody “inhibits interaction”between two proteins when the antibody disrupts, reduces, or completelyeliminates an interaction between the two proteins. An antibody of thepresent disclosure, or fragment thereof, “inhibits interaction” betweentwo proteins when the antibody or fragment thereof binds to one of thetwo proteins.

An “agonist” antibody or an “activating” antibody is an antibody, suchas an agonist anti-Sortilin antibody of the present disclosure, thatinduces (e.g., increases) one or more activities or functions of theantigen after the antibody binds the antigen.

A “blocking” antibody, an “antagonist” antibody, or an “inhibitory”antibody is an antibody, such as an anti-Sortilin antibody of thepresent disclosure, that inhibits or reduces (e.g., decreases) antigenbinding to one or more ligand after the antibody binds the antigen,and/or that inhibits or reduces (e.g., decreases) one or more activitiesor functions of the antigen after the antibody binds the antigen. Insome embodiments, blocking antibodies, antagonist antibodies, orinhibitory antibodies substantially or completely inhibit antigenbinding to one or more ligand and/or one or more activities or functionsof the antigen.

Antibody “effector functions” refer to those biological activitiesattributable to the Fc region (a native sequence Fc region or amino acidsequence variant Fc region) of an antibody, and vary with the antibodyisotype.

The term “Fc region” herein is used to define a C-terminal region of animmunoglobulin heavy chain, including native-sequence Fc regions andvariant Fc regions. Although the boundaries of the Fc region of animmunoglobulin heavy chain might vary, the human IgG heavy-chain Fcregion is usually defined to stretch from an amino acid residue atposition Cys226, or from Pro230, to the carboxyl-terminus thereof. TheC-terminal lysine (residue 447 according to the EU numbering system) ofthe Fc region may be removed, for example, during production orpurification of the antibody, or by recombinantly engineering thenucleic acid encoding a heavy chain of the antibody. Accordingly, acomposition of intact antibodies may comprise antibody populations withall K447 residues removed, antibody populations with no K447 residuesremoved, and antibody populations having a mixture of antibodies withand without the K447 residue. Suitable native-sequence Fc regions foruse in the antibodies of the present disclosure include human IgG1,IgG2, IgG3 and IgG4.

A “native sequence Fc region” comprises an amino acid sequence identicalto the amino acid sequence of an Fc region found in nature. Nativesequence human Fc regions include a native sequence human IgG1 Fc region(non-A and A allotypes); native sequence human IgG2 Fc region; nativesequence human IgG3 Fc region; and native sequence human IgG4 Fc regionas well as naturally occurring variants thereof.

A “variant Fc region” comprises an amino acid sequence which differsfrom that of a native sequence Fc region by virtue of at least one aminoacid modification, preferably one or more amino acid substitution(s).Preferably, the variant Fc region has at least one amino acidsubstitution compared to a native sequence Fc region or to the Fc regionof a parent polypeptide, e.g. from about one to about ten amino acidsubstitutions, and preferably from about one to about five amino acidsubstitutions in a native sequence Fc region or in the Fc region of theparent polypeptide. The variant Fc region herein will preferably possessat least about 80% homology with a native sequence Fc region and/or withan Fc region of a parent polypeptide, and most preferably at least about90% homology therewith, more preferably at least about 95% homologytherewith.

“Fc receptor” or “FcR” describes a receptor that binds to the Fc regionof an antibody. The preferred FcR is a native sequence human FcR.Moreover, a preferred FcR is one which binds an IgG antibody (a gammareceptor) and includes receptors of the FcγR1, FcγRII, and FcγRIIIsubclasses, including allelic variants and alternatively spliced formsof these receptors, FcγRII receptors include FcγRIIA (an “activatingreceptor”) and FcγRIIB (an “inhibiting receptor”), which have similaramino acid sequences that differ primarily in the cytoplasmic domainsthereof. Activating receptor FcγRIIA contains an immunoreceptortyrosine-based activation motif (“ITAM”) in its cytoplasmic domain.Inhibiting receptor FcγRIIB contains an immunoreceptor tyrosine-basedinhibition motif (“ITIM”) in its cytoplasmic domain. Other FcRs,including those to be identified in the future, are encompassed by theterm “FcR” herein. FcRs can also increase the serum half-life ofantibodies. As used herein, “percent (%) amino acid sequence identity”and “homology” with respect to a peptide, polypeptide or antibodysequence refers to the percentage of amino acid residues in a candidatesequence that are identical with the amino acid residues in the specificpeptide or polypeptide sequence, after aligning the sequences andintroducing gaps, if necessary, to achieve the maximum percent sequenceidentity, and not considering any conservative substitutions as part ofthe sequence identity. Alignment for purposes of determining percentamino acid sequence identity can be achieved in various ways that arewithin the skill in the art, for instance, using publicly availablecomputer software such as BLAST, BLAST-2, ALIGN or MEGALIGN™ (DNASTAR)software. Those skilled in the art can determine appropriate parametersfor measuring alignment, including any algorithms known in the artneeded to achieve maximal alignment over the full length of thesequences being compared.

An “isolated” cell is a molecule or a cell that is identified andseparated from at least one contaminant cell with which it is ordinarilyassociated in the environment in which it was produced. In someembodiments, the isolated cell is free of association with allcomponents associated with the production environment. The isolated cellis in a form other than in the form or setting in which it is found innature. Isolated cells are distinguished from cells existing naturallyin tissues, organs, or individuals. In some embodiments, the isolatedcell is a host cell of the present disclosure.

An “isolated” nucleic acid molecule encoding an antibody, such as ananti-Sortilin antibody of the present disclosure, is a nucleic acidmolecule that is identified and separated from at least one contaminantnucleic acid molecule with which it is ordinarily associated in theenvironment in which it was produced. Preferably, the isolated nucleicacid is free of association with all components associated with theproduction environment. The isolated nucleic acid molecules encoding thepolypeptides and antibodies herein is in a form other than in the formor setting in which it is found in nature. Isolated nucleic acidmolecules therefore are distinguished from nucleic acid encoding thepolypeptides and antibodies herein existing naturally in cells.

The term “vector,” as used herein, is intended to refer to a nucleicacid molecule capable of transporting another nucleic acid to which ithas been linked. One type of vector is a “plasmid,” which refers to acircular double stranded DNA into which additional DNA segments may beligated. Another type of vector is a phage vector. Another type ofvector is a viral vector, wherein additional DNA segments may be ligatedinto the viral genome. Certain vectors are capable of autonomousreplication in a host cell into which they are introduced (e.g.,bacterial vectors having a bacterial origin of replication and episomalmammalian vectors). Other vectors (e.g., non-episomal mammalian vectors)can be integrated into the genome of a host cell upon introduction intothe host cell, and thereby are replicated along with the host genome.Moreover, certain vectors are capable of directing the expression ofgenes to which they are operatively linked. Such vectors are referred toherein as “recombinant expression vectors,” or simply, “expressionvectors.” In general, expression vectors of utility in recombinant DNAtechniques are often in the form of plasmids. In the presentspecification, “plasmid” and “vector” may be used interchangeably as theplasmid is the most commonly used form of vector.

“Polynucleotide,” or “nucleic acid,” as used interchangeably herein,refer to polymers of nucleotides of any length, and include DNA and RNA.The nucleotides can be deoxyribonucleotides, ribonucleotides, modifiednucleotides or bases, and/or their analogs, or any substrate that can beincorporated into a polymer by DNA or RNA polymerase or by a syntheticreaction.

A “host cell” includes an individual cell or cell culture that can be orhas been a recipient for vector(s) for incorporation of polynucleotideinserts. Host cells include progeny of a single host cell, and theprogeny may not necessarily be completely identical (in morphology or ingenomic DNA complement) to the original parent cell due to natural,accidental, or deliberate mutation. A host cell includes cellstransfected in vivo with a polynucleotide(s) of the present disclosure.

“Carriers” as used herein include pharmaceutically acceptable carriers,excipients, or stabilizers that are nontoxic to the cell or mammal beingexposed thereto at the dosages and concentrations employed.

The term “about” as used herein refers to the usual error range for therespective value readily known to the skilled person in this technicalfield. Reference to “about” a value or parameter herein includes (anddescribes) embodiments that are directed to that value or parameter perse.

As used herein and in the appended claims, the singular forms “a,” “an,”and “the” include plural reference unless the context clearly indicatesotherwise. For example, reference to an “antibody” is a reference tofrom one to many antibodies, such as molar amounts, and includesequivalents thereof known to those skilled in the art, and so forth.

It is understood that aspect and embodiments of the present disclosuredescribed herein include “comprising,” “consisting,” and “consistingessentially of” aspects and embodiments.

Overview

The present disclosure relates, in part, to anti-Sortilin antibodiesthat exhibit one or more improved and/or enhanced functionalcharacteristics relative to an anti-Sortilin antibody, S-60, having aheavy chain variable region and a light chain variable region asdescribed in WO2016164637. Non-limiting improved and/or enhancedfunctional properties include, for example, antibodies capable ofbinding Sortilin with higher affinity, reducing cell surface levels ofSortilin, decreasing the half-maximal effective concentration (EC₅₀) toreduce cell surface levels of Sortilin, improving the maximal reductionof cell surface levels of Sortilin, increasing extracellular secretionof a Sortilin ligand: e.g Progranulin (PGRN), decreasing thehalf-maximal effective concentration (EC₅₀) to block PGRN binding toSortilin, improving the maximal blocking of PGRN binding to Sortilin, orany combination thereof. Also contemplated herein are anti-Sortilinantibodies with different Fc variants that exhibit one or more improvedand/or enhanced functional characteristics, including decreasing thehalf-maximal effective concentration (EC₅₀) to reduce cell surfacelevels of Sortilin, improving the maximal reduction of cell surfacelevels of Sortilin, increasing extracellular secretion of a Sortilinligand: e.g Progranulin (PGRN), decreasing the half-maximal effectiveconcentration (EC₅₀) to block PGRN binding to Sortilin, and improvingthe maximal blocking of PGRN binding to Sortilin.

In some embodiments, anti-Sortilin antibodies of the present disclosurehave higher potencies in reducing cell surface levels of Sortilinrelative to an anti-Sortilin antibody having a heavy chain variableregion and a light chain variable region corresponding to S-60. In someembodiments, potency is measured by half maximal effective concentration(EC₅₀) for reduction of cell surface cell levels of SORT1.

The present disclosure further relates to methods of making and usinganti-Sortilin antibodies as described herein; pharmaceuticalcompositions containing such antibodies; nucleic acids encoding suchantibodies; and host cells containing nucleic acids encoding suchantibodies.

In some embodiments, the anti-Sortilin antibodies of the presentdisclosure may have one or more activities that are due, at least inpart, to the ability of the antibodies to reduce levels (e.g., cellsurface levels) of Sortilin by inducing degradation, down regulation,cleavage, receptor desensitization, and/or lysosomal targeting ofSortilin. In some embodiments, the anti-Sortilin antibodies exhibit oneor more of the following properties: a. have a dissociation constant(K_(D)) for human Sortilin that is lower than that of an anti-Sortilinantibody having a heavy chain variable region and a light chain variableregion corresponding to S-60; b. decrease cell surface levels ofSortilin (e.g., decrease cell surface levels of Sortilin on engineeredcells expressing human Sortilin in vitro) with a half-maximal effectiveconcentration (EC₅₀) that is lower than that of an anti-Sortilinantibody having a heavy chain variable region and a light chain variableregion corresponding to S-60; c. have a dissociation constant (K_(D))for human Sortilin that may range from about 0.560 nM to about 1.63 nM,for example when the K_(D) is determined by fluorescent activated cellsorting (FACS); d. have a dissociation constant (K_(D)) for humanSortilin that may range from about 0.270 nM to about 2.910 nM, forexample when the K_(D) is determined by BioLayer Interferometry; e.decrease cell surface levels of Sortilin (e.g., decreases cell surfacelevels of Sortilin on engineered cells expressing human Sortilin invitro) with a half-maximal effective concentration (EC₅₀) that may rangefrom about 72.58 pM to about 103.6 pM, for example when the EC₅₀ isdetermined in vitro by FACS; f. reduce cell surface levels of Sortilinwith a maximum reduction that may range from about 51.33% to 88.76%; g.increase extracellular secretion of PGRN higher than that of ananti-Sortilin antibody having a heavy chain variable region and a lightchain variable region corresponding to S-60; h. block PGRN binding toSortilin (e.g., block binding of PGRN of Sortilin on engineered cellsexpressing human Sortilin in vitro) with a half-maximal effectiveconcentration (EC₅₀) that may range from about 0.325 nM to about 2.27nM, for example when the EC₅₀ is determined in vitro by FACS; and/or i.improve the maximal blocking of PGRN binding to Sortilin higher thanthat of an anti-Sortilin antibody having a heavy chain variable regionand a light chain variable region corresponding to S-60.

Advantageously, anti-Sortilin antibodies of the present disclosurereduce cell surface levels (e.g., up to approximately 3.02-fold) ofSortilin more potently (e.g., with a lower EC₅₀) as compared to acontrol anti-Sortilin antibody (e.g., a control anti-Sortilin antibodyhaving a heavy chain variable region and a light chain variable regioncorresponding to S-60) (See e.g., Example 2). In some embodiments,anti-Sortilin antibodies of the present disclosure reduce binding ofPGRN to Sortilin with a lower EC₅₀ (up to approximately 5.36-fold) morepotently as compared to a control anti-Sortilin antibody (e.g., acontrol anti-Sortilin antibody having a heavy chain variable region anda light chain variable region corresponding to S-60) (See e.g., Example3). Moreover, advantageously, anti-Sortilin antibodies of the presentdisclosure have a higher affinity (e.g., up to approximately 2.79-foldhigher affinity) for Sortilin (e.g., a lower K_(D) value as measured byFACS or BioLayer Interferometry) as compared to a control anti-Sortilinantibody (e.g., a control anti-Sortilin antibody having a heavy chainvariable region and a light chain variable region corresponding to S-60(See e.g., Examples 1 and 4). Surprisingly, higher affinity for Sortilindoes not necessarily correlate with an increase in ability or potency ofreduction of cell surface levels of Sortilin (See, e.g., Examples 2 and4) nor an increase in ability or potency of blocking binding of PGRN toSortilin (See, e.g., Examples 3 and 4).

The present disclosure further relates to anti-Sortilin antibodies withimproved stability during manufacturing, storage, and in vivoadministration. In some embodiments, anti-Sortilin antibodies of thepresent disclosure have improved stability under various stressconditions (See, e.g., Example 4).

Sortilin Proteins

In one aspect, the present disclosure provides antibodies, such asisolated (e.g., monoclonal) antibodies, that interact with or otherwisebind to a region, such as an epitope, within a Sortilin protein of thepresent disclosure. In some embodiments, the antibodies interact with orotherwise bind to a region, such as an epitope, within a Sortilinprotein of the present disclosure with improved/enhanced kinetics (e.g.,relative to an anti-Sortilin antibody having a heavy chain variableregion and a light chain variable region corresponding to S-60). In someembodiments, the antibodies interact with or otherwise bind to a region,such as an epitope, within a human Sortilin protein, with a half-maximaleffective concentration (EC₅₀) that is lower than that of a controlantibody (e.g., relative to an anti-Sortilin antibody having a heavychain variable region and a light chain variable region corresponding toS-60). In some embodiments, anti-Sortilin antibodies of the presentdisclosure bind to a Sortilin protein and modulate one or more Sortilinactivities after binding to the Sortilin protein, for example, anactivity associated with Sortilin expression on a cell. Sortilinproteins of the present disclosure include, without limitation, amammalian Sortilin protein, human Sortilin protein, mouse Sortilinprotein, and rat Sortilin protein. Exemplary Sortilin protein sequencesare shown in Table 32.

Sortilin is variously referred to as sortilin 1, SORT1, 100 kDa NTreceptor, glycoprotein 95 (GP95), Progranulin receptor (PGRN-R), andneurotensin receptor 3 (NT-3 or NTR-3). Sortilin is an 831 amino acidprotein that encodes a type I membrane receptor. Various Sortilinhomologs are known, including without limitation, human Sortilin, ratSortilin, and mouse Sortilin. The amino acid sequence of human Sortilinis set forth below as SEQ ID NO: 81 (with key amino acid residuespredicted to participate in Progranulin binding depicted in bold and,and the predicted pro-NGF binding region underlined):

        10         20         30         40MERPWGAADG LSRWPHGLGL LLLLQLLPPS TLSQDRLDAP        50         60         70         80PPPAAPLPRW SGPIGVSWGL RAAAAGGAFP RGGRWRRSAP        90        100        110        120GEDEECGRVR DFVAKLANNT HQHVFDDLRG SVSLSWVGDS       130        140        150        160TGVILVLTTF HVPLVIMTFG QSKLYRSEDY GKNFKDITDL       170        180        190        200INNTFIRTEF GMAIGPENSG KVVLTAEVSG GSRGGRIFRS       210        220        230        240SDFAKNFVQT DLPFHPLTQM MYSPQNSDYL LALSTENGLW       250        260        270        280VSKNFGGKWE EIHKAVCLAK WGSDNTIFFT TYANGSCKAD       290        300        310        320LGALELWRTS DLGKSFKTIG VKIYSFGLGG RFLFASVMAD       330        340        350        360KDTTRRIHVS TDQGDTWSMA QLPSVGQEQF YSILAANDDM       370        380        390        400VFMHVDEPGD TGFGTIFTSD DRGIVYSKSL DRHLYTTTGG       410        420        430        440ETDFTNVTSL RGVYITSVLS EDNSIQTMIT FDQGGRWTHL       450        460        470        480RKPENSECDA TAKNKNECSL HIHASYSISQ KLNVPMAPLS       490        500        510        520EPNAVGIVIA HGSVGDAISV MVPDVYISDD GGYSWTKMLE       530        540        550        560GPHYYTILDS GGIIVAIEHS SRPINVIKFS TDEGQCWQTY       570        580        590        600TFTRDPIYFT GLASEPGARS MNISIWGFTE SFLTSQWVSY       610        620        630        640TIDFKDILER NCEEKDYTIW LAHSTDPEDY EDGCILGYKE       650        660        670        680QFLRLRKSSV CQNGRDYVVT KQPSICLCSL EDFLCDFGYY       690        700        710        720RPENDSKCVE QPELKGHDLE FCLYGREEHL TTNGYRKIPG       730        740        750        760DKCQGGVNPV REVKDLKKKC TSNFLSPEKQ NSKSNSVPII       770        780        790        800LAIVGLMLVT VVAGVLIVKK YVCGGRFLVH RYSVLQQHAE       810        820        830 ANGVDGVDAL DTASHTNKSG YHDDSDEDLL E

The amino acid sequence of mouse Sortilin is set forth in SEQ ID NO: 82:

MERPRGAADG LLRWPLGLLL LLQLLPPAAV GQDRLDAPPP PAPPLLRWAG PVGVSWGLRAAAPGGPVPRA GRWRRGAPAE DQDCGRLPDF IAKLTNNTHQ HVFDDLSGSV SLSWVGDSTGVILVLTTFQV PLVIVSFGQS KLYRSEDYGK NFKDITNLIN NTFIRTEFGM AIGPENSGKVILTAEVSGGS RGGRVFRSSD FAKNFVQTDL PFHPLTQMMY SPQNSDYLLA LSTENGLWVSKNFGEKWEEI HKAVCLAKWG PNNIIFFTTH VNGSCKADLG ALELWRTSDL GKTFKTIGVKIYSFGLGGRF LFASVMADKD TTRRIHVSTD QGDTWSMAQL PSVGQEQFYS ILAANEDMVFMHVDEPGDTG FGTIFTSDDR GIVYSKSLDR HLYTTTGGET DFTNVTSLRG VYITSTLSEDNSIQSMITFD QGGRWEHLRK PENSKCDATA KNKNECSLHI HASYSISQKL NVPMAPLSEPNAVGIVIAHG SVGDAISVMV PDVYISDDGG YSWAKMLEGP HYYTILDSGG IIVAIEHSNRPINVIKFSTD EGQCWQSYVF TQEPIYFTGL ASEPGARSMN ISIWGFTESF ITRQWVSYTVDFKDILERNC EEDDYTTWLA HSTDPGDYKD GCILGYKEQF LRLRKSSVCQ NGRDYVVAKQPSVCPCSLED FLCDFGYFRP ENASECVEQP ELKGHELEFC LYGKEEHLTT NGYRKIPGDKCQGGMNPARE VKDLKKKCTS NFLNPTKQNS KSNSVPIILA IVGLMLVTVV AGVLIVKKYVCGGRFLVHRY SVLQQHAEAD GVEALDSTSH AKSGYHDDSD EDLLE

The amino acid sequence of rat Sortilin is set forth in SEQ ID NO: 83:

MERPRGAADG LLRWPLGLLL LLQLLPPAAV GQDRLDAPPP PAPPLLRWAG PVGVSWGLRAAAPGGPVPRA GRWRRGAPAE DQDCGRLPDF IAKLTNNTHQ HVFDDLSGSV SLSWVGDSTGVILVLTTFQV PLVIVSFGQS KLYRSEDYGK NFKDITNLIN NTFIRTEFGM AIGPENSGKVILTAEVSGGS RGGRVFRSSD FAKNFVQTDL PFHPLTQMMY SPQNSDYLLA LSTENGLWVSKNFGEKWEEI HKAVCLAKWG PNNIIFFTTH VNGSCKADLG ALELWRTSDL GKTFKTIGVKIYSFGLGGRF LFASVMADKD TTRRIHVSTD QGDTWSMAQL PSVGQEQFYS ILAANDDMVFMHVDEPGDTG FGTIFTSDDR GIVYSKSLDR HLYTTTGGET DFTNVTSLRG VYITSTLSEDNSIQSMITFD QGGRWEHLQK PENSKCDATA KNKNECSLHI HASYSISQKL NVPMAPLSEPNAVGIVIAHG SVGDAISVMV PDVYISDDGG YSWAKMLEGP HYYTILDSGG IIVAIEHSNRPINVIKFSTD EGQCWQSYVF SQEPVYFTGL ASEPGARSMN ISIWGFTESF LTRQWVSYTIDFKDILERNC EENDYTTWLA HSTDPGDYKD GCILGYKEQF LRLRKSSVCQ NGRDYVVAKQPSICPCSLED FLCDFGYFRP ENASECVEQP ELKGHELEFC LYGKEEHLTT NGYRKIPGDRCQGGMNPARE VKDLKKKCTS NFLNPKKQNS KSSSVPIILA IVGLMLVTVV AGVLIVKKYVCGGRFLVHRY SVLQQHAEAD GVEALDTASH AKSGYHDDSD EDLLE

In some embodiments, the Sortilin is a preprotein that includes a signalsequence. In some embodiments, the Sortilin is a mature protein. In someembodiments, the mature Sortilin protein does not include a signalsequence. In some embodiments, the mature Sortilin protein is expressedon a cell.

Sortilin proteins of the present disclosure include several domains,including without limitation, a signal sequence, a propeptide, a luminaldomain, a Vps10p domain, a 10 CC domain, a transmembrane domain and acytoplasmic domain. Additionally, proteins of the present disclosure areexpressed at high levels in a number of tissues, including withoutlimitation, the brain, spinal cord, heart and skeletal muscle, thyroid,placenta, and testis.

Accordingly, as used herein a “Sortilin” protein of the presentdisclosure includes, without limitation, a mammalian Sortilin protein,human Sortilin protein, primate Sortilin protein, mouse Sortilinprotein, and rat Sortilin protein. Additionally, anti-Sortilinantibodies of the present disclosure may bind an epitope within one ormore of a mammalian Sortilin protein, human Sortilin protein, primateSortilin, mouse Sortilin protein, and rat Sortilin protein.

Sortilin Protein Domains

Sortilin proteins of the present disclosure contain several domains,such as a Vps10p domain that contains an Asp-box motif, a ten-bladedbeta-propeller structure, and a hydrophobic loop; and a 10 CC domain.

As disclosed herein, interactions between Sortilin proteins of thepresent disclosure and pro-neurotrophins or neurotrophins are mediatedby the Vps10p domain that contains a ten-bladed beta-propeller structureand an Asp-box motif. In certain embodiments, Sortilin proteins of thepresent disclosure contain a Vps10p domain that includes a ten-bladedbeta-propeller structure and is located within amino acid residues78-611 of human Sortilin (SEQ ID NO: 81) or amino acid residues of amammalian Sortilin that correspond to amino acid residues 78-611 of SEQID NO: 81. In certain embodiments, amino acid residues 190-220 of humanSortilin (SEQ ID NO: 81) or amino acid residues of a mammalian Sortilinthat correspond to amino acid residues 190-220 of SEQ ID NO: 81 arelocated within the Vps10p domain.

Vps10p domains of the present disclosure may include an Asp-box motif.As used herein, Asp-box motifs have the following sequence:(S/T)-X-(D/N)-X-X-X-X-(W/F/Y) (SEQ ID NO: 84), orX-X-(S/T)-X-(D/N)-X-G-X-(T/S)-(W/F/Y)-X (SEQ ID NO: 85), where Xrepresents any amino acid. In human Sortilin, the Asp-box motif isSSDFAKNF (SEQ ID NO:86), located at amino acid residues 200-207 of humanSortilin. Accordingly, in certain embodiments, an Asp-box motif islocated at amino acid residues 200-207 of human Sortilin (SEQ ID NO: 81)or amino acid residues of a mammalian Sortilin that correspond to aminoacid residues 200-207 of SEQ ID NO: 81.

As disclosed herein, interactions between Sortilin proteins of thepresent disclosure and p75 are mediated by the 10CC domain of thehydrophobic loop of the Vps10p domain.

In certain embodiments, Sortilin proteins of the present disclosurecontain a 10CC domain that is located within amino acid residues 610-757of human Sortilin (SEQ ID NO: 81) or amino acid residues of a mammalianSortilin that correspond to amino acid residues 610-757 of SEQ ID NO:81. In preferred embodiments, amino acid residues 592-593, 610-660,and/or 667-749 of human Sortilin (SEQ ID NO: 81) or amino acid residuesof a mammalian Sortilin that correspond to amino acid residues 592-593,610-660, and/or 667-749 of SEQ ID NO: 81 are located within the 10CCdomain of Sortilin.

In other embodiments, Sortilin proteins of the present disclosurecontain a hydrophobic loop within the Vps10p domain that is locatedwithin amino acid residues 130-141 of human Sortilin (SEQ ID NO: 81) oramino acid residues of a mammalian Sortilin that correspond to aminoacid residues 130-141 of SEQ ID NO: 81.

As one of skill in the art will appreciate, the beginning and endingresidues of the domains of the present disclosure may vary dependingupon the computer modeling program used or the method used fordetermining the domain.

Sortilin Binding Partners

Sortilin proteins of the present disclosure can interact with (e.g.,bind to) one or more proteins including, without limitation, Progranulin(PGRN) protein; neurotrophins, such as pro-neurotrophins,pro-neurotrophin-3, neurotrophin-3, pro-neurotrophin-4/5,neurotrophin-4/5, pro-nerve growth factor (Pro-NGF), nerve growth factor(NGF), pro-brain-derived neurotrophic factor (Pro-BDNF), andbrain-derived neurotrophic factor (BDNF); neurotensin, p75, lipoproteinlipase (LpL), apolipoprotein AV (APOA5), apolipoprotein E (APOE),amyloid precursor protein, A beta peptide, PCSK9, p75NTR, and receptorassociated protein (RAP).

Progranulin (PGRN)

Sortilin proteins of the present disclosure have been shown to interact(e.g., bind to) directly with Progranulin and mediate the degradation ofProgranulin (e.g., Zheng, Y et al., (2011) PLoS ONE 6(6): e21023).

Progranulin is variously referred to as PGRN, proepithelin,granulin-epithelin precursor, PC (prostate cancer) cell-derived growthfactor (PCDGF), and acrogranin. Progranulin is a 593 amino acid proteinthat encodes a 68.5 kD a secreted glycoprotein that has 7.5 repeats ofsmaller granulin (epithelin) motifs, ranging from 6-25 kDa, which can beproteolytically cleaved from the precursor PGRN. Examples of Progranulincleavage products include, without limitation, granulin A/Epithelins 1,granulin B Epithelins 2, granulin C, granulins D, granulin E, granulinF, granulin G and any other known peptide products derived fromProgranulin.

Accordingly, anti-Sortilin antibodies of the present disclosure thatincrease Progranulin levels, decrease cell surface levels of Sortilin,and/or block the interaction (e.g., binding) between Sortilin andProgranulin would be beneficial for preventing, lowering the risk of, ortreating conditions and/or diseases associated with decreased levels ofProgranulin expression and/or activity, cell death (e.g., neuronal celldeath), frontotemporal dementia, Alzheimer's disease, vascular dementia,seizures, retinal dystrophy, a traumatic brain injury, a spinal cordinjury, long-term depression, atherosclerotic vascular diseases,undesirable symptoms of normal aging, dementia, mixed dementia,Creutzfeldt-Jakob disease, normal pressure hydrocephalus, amyotrophiclateral sclerosis, Huntington's disease, taupathy disease, stroke, acutetrauma, chronic trauma, lupus, acute and chronic colitis, Crohn'sdisease, inflammatory bowel disease, ulcerative colitis, malaria,essential tremor, central nervous system lupus, Behcet's disease,Parkinson's disease, dementia with Lewy bodies, multiple system atrophy,intervertebral disc degeneration, Shy-Drager syndrome, progressivesupranuclear palsy, cortical basal ganglionic degeneration, acutedisseminated encephalomyelitis, granulomartous disorders, Sarcoidosis,diseases of aging, age related macular degeneration, glaucoma, retinitispigmentosa, retinal degeneration, respiratory tract infection, sepsis,eye infection, systemic infection, inflammatory disorders, arthritis,multiple sclerosis, metabolic disorder, obesity, insulin resistance,type 2 diabetes, tissue or vascular damage, an injury, and/or one ormore undesirable symptoms of normal aging. Additionally, anti-Sortilinantibodies of the present disclosure that increase Progranulin levels,decrease cell surface levels of Sortilin, and/or block the interaction(e.g., binding) between Sortilin and Progranulin may inhibit interactionbetween Sortilin and Progranulin, may induce one or more Progranulinactivities, may reduce the endosomal internalization of Progranulin, orfragments thereof, and/or may increase the effective concentration ofProgranulin.

In some embodiments, anti-Sortilin antibodies of the present disclosurethat increase Progranulin levels, decrease cell surface levels ofSortilin, and/or block the interaction (e.g., binding) between Sortilinand Progranulin bind to one or more amino acids within amino acidresidues 131-138, 175-181, 190-220, 199-220, 190-211, 196-207, 196-199,200-207, 203-207, 207-231, 207-227, 212-221, 233-243, 237-247, 237-260,297-317, 314-338, 367-391, 429-443, 623-632, and/or 740-749 of humanSortilin (SEQ ID NO: 81); or within amino acid residues of a mammalianSortilin that corresponds to amino acid residues 131-138, 175-181,190-220, 199-220, 190-211, 196-207, 196-199, 200-207, 203-207, 207-231,207-227, 212-221, 233-243, 237-247, 237-260, 297-317, 314-338, 367-391,429-443, 623-632, and/or 740-749 of SEQ ID NO: 81. In other embodiments,anti-Sortilin antibodies of the present disclosure that increaseProgranulin levels, decrease cell surface levels of Sortilin, and/orblock the interaction (e.g., binding) between Sortilin and Progranulinmay bind one or more amino acids of amino acid residues His131, Val132,Pro133, Leu134, Val135, Ile136, Met137, Thr138, Arg196, Phe198, Arg199,Phe203, Lys205, Phe207, Thr210, Thr218, Tyr222, Ser223, Ser227, Ser242,Lys243, Lys248, Lys254, Lys260, Ser305, Phe306, Gly307, Arg311, Phe314,Ser316, Arg325, Arg326, Ile327, Phe350, Tyr351, Ser352, Ile353, Asn373,Ser379, Arg382, Tyr386, Ser595, and/or Glu700 of human Sortilin (SEQ IDNO: 81); or of amino acid residues of a mammalian Sortilin thatcorresponds to one or more amino acid residues His131, Val132, Pro133,Leu134, Val135, Ile136, Met137, Thr138, Arg196, Phe198, Arg199, Phe203,Lys205, Phe207, Thr210, Thr218, Tyr222, Ser223, Ser227, Ser242, Lys243,Lys248, Lys254, Lys260, Ser305, Phe306, Gly307, Arg311, Phe314, Ser316,Arg325, Arg326, Ile327, Phe350, Tyr351, Ser352, Ile353, Asn373, Ser379,Arg382, Tyr386, Ser595, and/or Glu700 of SEQ ID NO: 81.

Other Sortilin Ligands

Sortilin proteins of the present disclosure have been shown to interact(e.g., bind) directly with pro-neurotrophins (e.g., pro-NGF), whichharbor a pro-domain and are typically pro-apoptotic. This binding may bemediated through a linear epitope on Sortilin that corresponds to aminoacid residues 163-174 of SEQ ID NO: 81. Sortilin proteins of the presentdisclosure also have been shown to interact (e.g., bind) withneurotensin within the beta-propeller structure of Sortilin, and animportant contact has been shown to at serine 283 of human Sortilin.

Sortilin proteins of the present disclosure have also been shown tointeract (e.g., bind) with the low affinity nerve growth factor (NGF)receptor (p75) within the 10CC domain of Sortilin or the hydrophobicloop of the Vps10p domain of Sortilin. As disclosed herein, Sortilinproteins of the present disclosure can function as a co-receptor withp75 for pro-neurotrophins, which induce apoptotic signaling. Sortilinproteins of the present disclosure have further been shown to interact(e.g., bind) with amyloid precursor protein (APP).

Sortilin proteins of the present disclosure have been shown to interact(e.g., bind) with the lipoprotein lipase (LpL). As disclosed herein,Sortilin proteins of the present disclosure bind to and modify thedegradation of LpL. Sortilin proteins of the present disclosure havebeen shown to interact (e.g., bind) with the apolipoprotein AV (APOA5).As disclosed herein, Sortilin proteins of the present disclosure bind toand modify the degradation of APOA5.

Sortilin proteins of the present disclosure have been shown to interact(e.g., bind) with the apolipoprotein E (APOE, APOE2, APOE3, APOE4). Asdisclosed herein, Sortilin proteins of the present disclosure bind toand modify the degradation and transport of APOE as well as agents thatAPOE carries such as the A beta peptide. In addition, Sortilin proteinsof the present disclosure have been shown to interact (e.g., bind) withreceptor-associate protein (RAP). Further, Sortilin proteins of thepresent disclosure have been shown to interact (e.g., bind) withProprotein convertase subtilisin/kexin type 9 (PCSK9), and secrete it tothe circulation.

In some embodiments of any of the anti-Sortilin antibodies,anti-Sortilin antibodies of the present disclosure may also inhibit(e.g., block) the interaction between Sortilin and one or more otherSortilin ligands of the present disclosure. In some embodiments, the oneor more other ligands is one or more of pro-neurotrophins, neurotensin,low affinity nerve growth factor (NGF) receptor (p75), amyloid precursorprotein, lipoprotein lipase, apolipoprotein AV, apolipoprotein,receptor-associated protein, and/or proprotein convertasesubtilisin/kexin type 9. Such antibodies may be beneficial forpreventing, lowering the risk of, or treating conditions and/or diseasesassociated with decreased levels of one or more other Sortilin ligandsexpression and/or activity, cell death (e.g., neuronal cell death). Insome embodiments, the one or more other ligands is one or more ofpro-neurotrophins, neurotensin, low affinity nerve growth factor (NGF)receptor (p75), amyloid precursor protein, lipoprotein lipase,apolipoprotein AV, apolipoprotein, receptor-associated protein, and/orproprotein convertase subtilisin/kexin type 9.

Anti-Sortilin antibodies of the present disclosure that inhibit (e.g.,block) the interaction between Sortilin and one or more other Sortilinligands of the present disclosure may also prevent cell death (e.g.,apoptosis) induced by one or more other Sortilin ligands. In someembodiments, the one or more other ligands is one or more ofpro-neurotrophins, neurotensin, low affinity nerve growth factor (NGF)receptor (p75), amyloid precursor protein, lipoprotein lipase,apolipoprotein AV, apolipoprotein, receptor-associated protein, and/orproprotein convertase subtilisin/kexin type 9.

In some embodiments of any of the anti-Sortilin antibodies,anti-Sortilin antibodies of the present disclosure that inhibit (e.g.,block) the interaction between Sortilin and one or more Sortilin ligandsof the present disclosure bind one or more amino acids within amino acidresidues within amino acid residues 131-138, 175-181, 190-220, 199-220,190-211, 196-207, 196-199, 200-207, 203-207, 207-231, 207-227, 212-221,233-243, 237-247, 237-260, 297-317, 314-338, 367-391, 429-443, 623-632,and/or 740-749 of human Sortilin (SEQ ID NO: 81); or within amino acidresidues of a mammalian Sortilin that corresponds to amino acid residues131-138, 175-181, 190-220, 199-220, 190-211, 196-207, 196-199, 200-207,203-207, 207-231, 207-227, 212-221, 233-243, 237-247, 237-260, 297-317,314-338, 367-391, 429-443, 623-632, and/or 740-749 of SEQ ID NO: 81. Insome embodiments, the one or more other ligands is one or more ofpro-neurotrophins, neurotensin, low affinity nerve growth factor (NGF)receptor (p75), amyloid precursor protein, lipoprotein lipase,apolipoprotein AV, apolipoprotein, receptor-associated protein, and/orproprotein convertase subtilisin/kexin type 9.

In other embodiments of any of the anti-Sortilin antibodies,anti-Sortilin antibodies of the present disclosure that inhibit (e.g.,block) the interaction between Sortilin and one or more Sortilin ligandsof the present disclosure bind one or more amino acids of amino acidresidues His131, Val132, Pro133, Leu134, Val135, Ile136, Met137, Thr138,Arg196, Phe198, Arg199, Phe203, Lys205, Phe207, Thr210, Thr218, Tyr222,Ser223, Ser227, Ser242, Lys243, Lys248, Lys254, Lys260, Ser305, Phe306,Gly307, Arg311, Phe314, Ser316, Arg325, Arg326, Ile327, Phe350, Tyr351,Ser352, Ile353, Asn373, Ser379, Arg382, Tyr386, Ser595, and/or Glu700 ofhuman Sortilin (SEQ ID NO: 81); or of amino acid residues of a mammalianSortilin that corresponds to one or more amino acid residues His131,Val132, Pro133, Leu134, Val135, Ile136, Met137, Thr138, Arg196, Phe198,Arg199, Phe203, Lys205, Phe207, Thr210, Thr218, Tyr222, Ser223, Ser227,Ser242, Lys243, Lys248, Lys254, Lys260, Ser305, Phe306, Gly307, Arg311,Phe314, Ser316, Arg325, Arg326, Ile327, Phe350, Tyr351, Ser352, Ile353,Asn373, Ser379, Arg382, Tyr386, Ser595, and/or Glu700 of SEQ ID NO: 81.In some embodiments, the one or more other ligands is one or more ofpro-neurotrophins, neurotensin, low affinity nerve growth factor (NGF)receptor (p75), amyloid precursor protein, lipoprotein lipase,apolipoprotein AV, apolipoprotein, receptor-associated protein, and/orproprotein convertase subtilisin/kexin type 9.

Sortilin Antibodies

Certain aspects of the present disclosure relate to anti-Sortilinantibodies comprising one or more improved and/or enhanced functionalcharacteristics. In some embodiments, anti-Sortilin antibodies of thepresent disclosure comprise one or more improved and/or enhancedfunctional characteristics relative to an anti-Sortilin antibody, S-60,having a heavy chain variable region and a light chain variable regionas described in WO2016164637. In some embodiments, anti-Sortilinantibodies of the present disclosure have an affinity for Sortilin(e.g., human Sortilin) that is higher than that of a controlanti-Sortilin antibody (e.g., a control anti-Sortilin antibodycomprising a heavy chain variable region and a light chain variableregion corresponding to S-60). In some embodiments, anti-Sortilinantibodies of the present disclosure decrease cellular levels (e.g.,cell surface levels) of Sortilin to a greater degree and with ahalf-maximal effective concentration (EC₅₀) that is lower than that of acontrol antibody (e.g., a control anti-Sortilin antibody comprising aheavy chain variable region and a light chain variable regioncorresponding to S-60). In some embodiments, anti-Sortilin antibodies ofthe present disclosure improve the maximal reduction of cell surfacelevels of Sortilin relative to an anti-Sortilin antibody comprising aheavy chain variable region and a light chain variable regioncorresponding to S-60. In some embodiments, anti-Sortilin antibodies ofthe present disclosure increase the secretion of extracellularProgranulin (PGRN) relative to an anti-Sortilin antibody comprising aheavy chain variable region and a light chain variable regioncorresponding to S-60. In some embodiments, anti-Sortilin antibodies ofthe present disclosure blocking binding of PGRN to Sortilin to a greaterdegree and with a half-maximal effective concentration (EC₅₀) that islower than that of a control antibody (e.g., a control anti-Sortilinantibody comprising a heavy chain variable region and a light chainvariable region corresponding to S-60). In some embodiments,anti-Sortilin antibodies of the present disclosure improve the maximalblocking of PGRN binding to Sortilin relative to an anti-Sortilinantibody comprising a heavy chain variable region and a light chainvariable region corresponding to S-60.

Also contemplated herein are anti-Sortilin antibodies with different Fcvariants that exhibit one or more improved and/or enhanced functionalcharacteristics relative to an anti-Sortilin antibody comprising a heavychain variable region and a light chain variable region corresponding toS-60, including decreasing the half-maximal effective concentration(EC₅₀) to reduce cell surface levels of Sortilin, improving the maximalreduction of cell surface levels of Sortilin, increasing extracellularsecretion of PGRN, decreasing the half-maximal effective concentration(EC₅₀) to block PGRN binding to Sortilin, and improving the maximalblocking of PGRN binding to Sortilin.

In some embodiments, an anti-Sortilin antibody of the present disclosureis a human antibody, a bispecific antibody, a monoclonal antibody, amultivalent antibody, a conjugated antibody, or a chimeric antibody

In a preferred embodiment, an anti-Sortilin antibody of the presentdisclosure is a monoclonal antibody.

Anti-Sortilin Antibody Binding Epitope

In some embodiments, anti-Sortilin antibodies of the present disclosurebind to a Sortilin protein of the present disclosure and/or naturallyoccurring variants. In some embodiments, anti-Sortilin antibodies of thepresent disclosure bind to a Sortilin protein, wherein the Sortilinprotein is a human protein. In some embodiments, anti-Sortilinantibodies of the present disclosure bind to a Sortilin protein, whereinthe Sortilin protein is a wild-type protein. In some embodiments,anti-Sortilin antibodies of the present disclosure bind to a Sortilinprotein, wherein the Sortilin protein is a naturally occurring variant.

In certain preferred embodiments, anti-Sortilin antibodies of thepresent disclosure bind specifically to a human Sortilin protein.

Certain aspects of the present disclosure provide anti-Sortilinantibodies that bind a discontinuous Sortilin epitope. In someembodiments, the discontinuous Sortilin epitope comprises two or morepeptides, three or more peptides, four or more peptides, five or morepeptides, six or more peptides, seven or more peptide, eight or morepeptides, nine or more peptides, or 10 or more peptides. In someembodiments, each of the peptides comprise five or more, six or more,seven or more, eight or more, nine or more, 10 or more, 11 or more, 12or more, 13 or more 14 or more, 15 or more, 16 or more, 17 or more, 18or more, 19 or more, 20 or more, 21 or more, 22 or more, 23 or more, 24or more, 25 or more, 26 or more, 27 or more, 28 or more, 29 or more, or30 or more amino acid residues of the amino acid sequence of SEQ ID NO:81; or five or more, six or more, seven or more, eight or more, nine ormore, 10 or more, 11 or more, 12 or more, 13 or more 14 or more, 15 ormore, 16 or more, 17 or more, 18 or more, 19 or more, 20 or more, 21 ormore, 22 or more, 23 or more, 24 or more, 25 or more, 26 or more, 27 ormore, 28 or more, 29 or more, or 30 or more amino acid residues on amammalian Sortilin protein corresponding to the amino acid sequence ofSEQ ID NO: 81. Other aspects of the present disclosure provideanti-Sortilin antibodies that bind to a conformational epitope ofSortilin.

Certain aspects of the present disclosure provide anti-Sortilinantibodies that bind to one or more amino acids within amino acidresidues 207-231 of human Sortilin (SEQ ID NO: 81); or within amino acidresidues on a mammalian Sortilin corresponding to amino acid residues207-231 of SEQ ID NO: 81. In some embodiments, the anti-Sortilinantibodies bind to one or more amino acids of amino acid residuesThr218, Tyr222, Ser223, and/or Ser227 of human Sortilin (SEQ ID NO: 81);or of amino acid residues on a mammalian Sortilin corresponding to aminoacid residues Thr218, Tyr222, Ser223, and/or Ser227 of SEQ ID NO: 81. Insome embodiments, the anti-Sortilin antibodies bind to one or more aminoacids of amino acid residues F105, L108, R109, G110, 1537, F569, E590,F592, L593, S595, and/or W597 of human Sortilin (SEQ ID NO: 81); or ofamino acid residues on a mammalian Sortilin corresponding to amino acidresidues F105, L108, R109, G110, 1537, F569, E590, F592, L593, S595,and/or W597 of SEQ ID NO: 81. Other aspects of the present disclosureprovide anti-Sortilin antibodies that bind to an epitope having aminoacid residues (S/T)-X-(D/N)-X-X-X-X-(W/F/Y), where X represents anyamino acid (SEQ ID NO: 84).

In some embodiments, anti-Sortilin antibodies of the present disclosurebind to a Sortilin protein of the present disclosure expressed on thesurface of cell and the naked antibodies inhibit interaction (e.g.,binding) between the Sortilin protein and a protein selected fromProgranulin (PGRN), a pro-neurotrophin, a neurotrophin,pro-neurotrophin-3, neurotrophin-3, pro-neurotrophin-4/5,neurotrophin-4/5, pro-nerve growth factor (pro-NGF), nerve growth factor(NGF), pro-brain-derived neurotrophic factor (pro-BDNF), brain-derivedneurotrophic factor (BDNF), neurotensin, p75, Sortilin propeptide(Sort-pro), amyloid precursor protein (APP), the A beta peptide,lipoprotein lipase (LpL), apolipoprotein AV (APOA5), apolipoprotein E(APOE), PCSK9, and receptor associated protein (RAP). In someembodiments, anti-Sortilin antibodies of the present disclosure bind toa Sortilin protein of the present disclosure expressed on the surface ofcell and the naked antibodies inhibit interaction (e.g., binding)between the Sortilin protein and PGRN.

In some embodiments, anti-Sortilin antibodies of the present disclosurethat bind to a Sortilin protein of the present disclosure inhibitinteraction (e.g., binding) between the Sortilin protein andProgranulin, a pro-neurotrophin, a neurotrophin, pro-neurotrophin-3,neurotrophin-3, pro-neurotrophin-4/5, neurotrophin-4/5, pro-nerve growthfactor (pro-NGF), nerve growth factor (NGF), pro-brain-derivedneurotrophic factor (pro-BDNF), brain-derived neurotrophic factor(BDNF), neurotensin, p75, Sortilin propeptide (Sort-pro), amyloidprecursor protein (APP), the A beta peptide, lipoprotein lipase (LpL),apolipoprotein AV (APOA5), apolipoprotein E (APOE), PCSK9, and receptorassociated protein (RAP) by reducing the effective levels of Sortilinthat is available to interact with these proteins either on the cellsurface or inside the cell.

In some embodiments, anti-Sortilin antibodies of the present disclosurethat bind to a Sortilin protein of the present disclosure inhibitinteraction (e.g., binding) between the Sortilin protein andProgranulin, a pro-neurotrophin, a neurotrophin, pro-neurotrophin-3,neurotrophin-3, pro-neurotrophin-4/5, neurotrophin-4/5, pro-nerve growthfactor (pro-NGF), nerve growth factor (NGF), pro-brain-derivedneurotrophic factor (pro-BDNF), brain-derived neurotrophic factor(BDNF), neurotensin, p75, Sortilin propeptide (Sort-pro), amyloidprecursor protein (APP), the A beta peptide, lipoprotein lipase (LpL),apolipoprotein AV (APOA5), apolipoprotein E (APOE), PCSK9, and receptorassociated protein (RAP) by inducing degradation of Sortilin.

In some embodiments, anti-Sortilin antibodies of the present disclosurebind to a human Sortilin, or a homolog thereof, including withoutlimitation, a mammalian Sortilin protein, or a non-human primateSortilin protein. In some embodiments, anti-Sortilin antibodies of thepresent disclosure specifically bind to human Sortilin. In someembodiments, anti-Sortilin antibodies of the present disclosure bind tohuman Sortilin and are not cross-reactive with Sortilin orthologs orhomologs from other species.

Anti-Sortilin Antibody Competitive Binding

In some embodiments, anti-Sortilin antibodies of the present disclosurecompetitively inhibit binding of at least one other anti-Sortilinantibody selected from any of the antibodies listed in Tables 1-29. Insome embodiments, anti-Sortilin antibodies of the present disclosurecompetitively inhibit binding of at least one antibody selected from:S-60, S-60-1, S-60-2, S-60-3, S-60-4, S-60-5, S-60-6, S-60-7, S-60-8,and S-60-9 (described in WO2016164637); or S-60-10, S-60-11, S-60-12,S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2[N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6[N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10[N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I],S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17[N33L], S-60-16; S-60-18, S-60-19, S-60-24; and any combination thereof.

In some embodiments, an anti-Sortilin antibody of the present disclosurecompetes with one or more anti-Sortilin antibodies selected from: S-60,S-60-1, S-60-2, S-60-3, S-60-4, S-60-5, S-60-6, S-60-7, S-60-8, andS-60-9 (described in WO2016164637); or S-60-10, S-60-11, S-60-12,S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2[N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6[N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10[N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I],S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17[N33L], S-60-16; S-60-18, S-60-19, S-60-24 and any combination thereof,for binding to Sortilin when the anti-Sortilin antibody reduces thebinding of one or more antibodies selected from: S-60, S-60-1, S-60-2,S-60-3, S-60-4, S-60-5, S-60-6, S-60-7, S-60-8, and S-60-9 (described inWO2016164637); or S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15[N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G],S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K],S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11[N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V],S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16;S-60-18, S-60-19, S-60-24; and any combination thereof to Sortilin by anamount the ranges from about 50% to 100%, as compared to binding toSortilin in the absence of the anti-Sortilin antibody.

In some embodiments, an anti-Sortilin antibody of the present disclosurecompetes with one or more anti-Sortilin antibodies selected from: S-60,S-60-1, S-60-2, S-60-3, S-60-4, S-60-5, S-60-6, S-60-7, S-60-8, andS-60-9 (described in WO2016164637); or S-60-10, S-60-11, S-60-12,S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2[N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6[N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10[N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I],S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17[N33L], S-60-16; S-60-18, S-60-19, S-60-24, and any combination thereof,for binding to Sortilin when the anti-Sortilin antibody reduces thebinding of one or more antibodies selected from: S-60, S-60-1, S-60-2,S-60-3, S-60-4, S-60-5, S-60-6, S-60-7, S-60-8, and S-60-9 (described inWO2016164637); or S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15[N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G],S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K],S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11[N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V],S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16;S-60-18, S-60-19, S-60-24; and any combination thereof to Sortilin by atleast 50%, at least 55%, by at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 95%, or100%, as compared to binding to Sortilin in the absence of theanti-Sortilin antibody.

In some embodiments, an anti-Sortilin antibody of the present disclosurethat reduces the binding of one or more antibodies selected from: S-60,S-60-1, S-60-2, S-60-3, S-60-4, S-60-5, S-60-6, S-60-7, S-60-8, andS-60-9 (described in WO2016164637); or S-60-10, S-60-11, S-60-12,S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2[N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6[N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10[N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I],S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17[N33L], S-60-16; S-60-18, S-60-19, S-60-24, and any combination thereof,to Sortilin by 100% indicates that the anti-Sortilin antibody essentialcompletely blocks the binding of one or more antibodies selected fromS-60, S-60-1, S-60-2, S-60-3, S-60-4, S-60-5, S-60-6, S-60-7, S-60-8,and S-60-9 (described in WO2016164637); or S-60-10, S-60-11, S-60-12,S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2[N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6[N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10[N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I],S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17[N33L], S-60-16; S-60-18, S-60-19, S-60-24, and any combination thereof,to Sortilin. In some embodiments, the anti-Sortilin antibody and the oneor more antibodies selected from S-60, S-60-1, S-60-2, S-60-3, S-60-4,S-60-5, S-60-6, S-60-7, S-60-8, and S-60-9 (described in WO2016164637);or S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)],S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R],S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q],S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12[N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A],S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19,S-60-24, and any combination thereof, are present in an amount thatcorresponds to a 10:1 ratio, 9:1 ratio, 8:1 ratio, 7:1 ratio, 6:1 ratio,5:1 ratio, 4:1 ratio, 3:1 ratio, 2:1 ratio, 1:1 ratio, 0.75:1 ratio,0.5:1 ratio, 0.25:1 ratio, 0.1:1 ratio, 0.075:1 ratio, 0.050:1 ratio,0.025:1 ratio, 0.01:1 ratio, 0.0075: ratio, 0.0050:1 ratio, 0.0025:1ratio, 0.001: ratio, 0.00075:1 ratio, 0.00050:1 ratio, 0.00025:1 ratio,0.0001: ratio, 1:10 ratio, 1:9 ratio, 1:8 ratio, 1:7 ratio, 1:6 ratio,1:5 ratio, 1:4 ratio, 1:3 ratio, 1:2 ratio, 1:0.75 ratio, 1:0.5 ratio,1:0.25 ratio, 1:0.1 ratio, 1:0.075 ratio, 1:0.050 ratio, 1:0.025 ratio,1:0.01 ratio, 1:0.0075 ratio, 1:0.0050 ratio, 1:0.0025 ratio, 1:0.001ratio, 1:0.00075 ratio, 1:0.00050 ratio, 1:0.00025 ratio, or 1:0.0001ratio of anti-Sortilin antibody to one or more antibodies selected fromS-60, S-60-1, S-60-2, S-60-3, S-60-4, S-60-5, S-60-6, S-60-7, S-60-8,and S-60-9 (described in WO2016164637); or S-60-10, S-60-11, S-60-12,S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2[N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6[N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10[N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I],S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17[N33L], S-60-16; S-60-18, S-60-19, S-60-24, and any combination thereof.In some embodiments, the anti-Sortilin antibody is present in excess byan amount that ranges from about 1.5-fold to 100-fold, or greater than100-fold compared to the amount of the one or more antibodies selectedfrom S-60, S-60-1, S-60-2, S-60-3, S-60-4, S-60-5, S-60-6, S-60-7,S-60-8, and S-60-9 (described in WO2016164637); or S-60-10, S-60-11,S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T],S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D],S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y],S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13[N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M],S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, S-60-24, and anycombination thereof. In some embodiments, the anti-Sortilin antibody ispresent in an amount that is about a 2-fold, 3-fold, 4-fold, 5-fold,6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 25-fold,30-fold, 35-fold, 40-fold, 45-fold, 50-fold, 55-fold, 60-fold, 65-fold,70-fold, 75-fold, 80-fold, 85-fold, 90-fold, 95-fold, or 100-fold excesscompared to the amount of the one or more antibodies selected from S-60,S-60-1, S-60-2, S-60-3, S-60-4, S-60-5, S-60-6, S-60-7, S-60-8, andS-60-9 (described in WO2016164637); or S-60-10, S-60-11, S-60-12,S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2[N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6[N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10[N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I],S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17[N33L], S-60-16; S-60-18, S-60-19, S-60-24, and any combination thereof.

In some embodiments, anti-Sortilin antibodies of the present disclosurebind to an epitope of human Sortilin that is the same as or overlapswith the Sortilin epitope bound by at least one antibody selected fromany of the antibodies listed in Tables 1-29. In some embodiments,anti-Sortilin antibodies of the present disclosure bind to an epitope ofhuman Sortilin that is the same as or overlaps with the Sortilin epitopebound by at least one antibody selected from: S-60, S-60-1, S-60-2,S-60-3, S-60-4, S-60-5, S-60-6, S-60-7, S-60-8, and S-60-9 (described inWO2016164637); or S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15[N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G],S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K],S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11[N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V],S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16;S-60-18, S-60-19, S-60-24; and any combination thereof. In someembodiments, anti-Sortilin antibodies of the present disclosure bindessentially the same Sortilin epitope bound by at least one antibodyselected from any of the antibodies listed in Tables 1-29. In someembodiments, anti-Sortilin antibodies of the present disclosure bindessentially the same Sortilin epitope bound by at least one antibodyselected from: S-60, S-60-1, S-60-2, S-60-3, S-60-4, S-60-5, S-60-6,S-60-7, S-60-8, and S-60-9 (described in WO2016164637); or S-60-10,S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1[N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5[N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9[N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F],S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16[N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, S-60-24; and anycombination thereof. Detailed exemplary methods for mapping an epitopeto which an antibody binds are provided in Morris (1996) “EpitopeMapping Protocols,” in Methods in Molecular Biology vol. 66 (HumanaPress, Totowa, N.J.).

Any suitable competition assay or Sortilin binding assay known in theart, such as BIAcore analysis, ELISA assays, or flow cytometry, may beutilized to determine whether an anti-Sortilin antibody competes withone or more antibodies selected from: S-60, S-60-1, S-60-2, S-60-3,S-60-4, S-60-5, S-60-6, S-60-7, S-60-8, and S-60-9 (described inWO2016164637); or S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15[N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G],S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K],S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11[N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V],S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16;S-60-18, S-60-19, S-60-24; and any combination thereof for binding toSortilin. In an exemplary competition assay, immobilized Sortilin orcells expressing Sortilin on the cell surface are incubated in asolution comprising a first labeled antibody that binds to Sortilin(e.g., human or non-human primate) and a second unlabeled antibody thatis being tested for its ability to compete with the first antibody forbinding to Sortilin. The second antibody may be present in a hybridomasupernatant. As a control, immobilized Sortilin or cells expressingSortilin is incubated in a solution comprising the first labeledantibody but not the second unlabeled antibody. After incubation underconditions permissive for binding of the first antibody to Sortilin,excess unbound antibody is removed, and the amount of label associatedwith immobilized Sortilin or cells expressing Sortilin is measured. Ifthe amount of label associated with immobilized Sortilin or cellsexpressing Sortilin is substantially reduced in the test sample relativeto the control sample, then that indicates that the second antibody iscompeting with the first antibody for binding to Sortilin. See, Harlowand Lane (1988) Antibodies: A Laboratory Manual ch. 14 (Cold SpringHarbor Laboratory, Cold Spring Harbor, N.Y.).

In some embodiments, an anti-Sortilin antibody of the present disclosurecompetes with an antibody comprising the heavy chain variable domain andthe light chain variable domain of an antibody selected from the groupconsisting of S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33(wt)], S-60-15.1 [N33T], S-60-16, S-60-18, S-60-19, S-60-24, and anycombination thereof for binding to Sortilin.

In some embodiments, an anti-Sortilin antibody of the present disclosurebinds essentially the same Sortilin epitope as an antibody comprisingthe heavy chain variable domain and the light chain variable domain ofan antibody selected from the group consisting of S-60-10, S-60-11,S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T],S-60-16, S-60-18, S-60-19, and S-60-24.

Additional anti-Sortilin antibodies, e.g., antibodies that specificallybind to a Sortilin protein of the present disclosure, may be identified,screened, and/or characterized for their physical/chemical propertiesand/or biological activities by various assays known in the art.

Anti-Sortilin Antibody Heavy Chain and Light Chain Variable Regions

A. Heavy Chain HVRs

In some embodiments, anti-Sortilin antibodies of the present disclosureinclude a heavy chain variable region comprising one or more (e.g., oneor more, two or more, or all three) HVRs selected from HVR-H1, HVR-H2,and HVR-H3 (as shown in Tables 14-16). In some embodiments, the heavychain variable region comprises an HVR-H1, an HVR-H2, and an HVR-H3 (asshown in Tables 14-16).

In some embodiments, the HVR-H1 comprises a sequence of YSISSGYYWG (SEQID NO: 1). In some embodiments, the HVR-H2 comprises a sequenceaccording to Formula I: TIYHSGSTYYNPSLX₁S (SEQ ID NO: 4), wherein X₁ isK or E. In some embodiments, the HVR-H2 comprises a sequence selectedfrom SEQ ID NOs: 2-3. In some embodiments, the HVR-H3 comprises asequence according to Formula II: ARQGSIX₁QGYYGMDV (SEQ ID NO: 7). Insome embodiments, the HVR-H3 comprises a sequence selected from SEQ IDNOs: 5-6.

In some embodiments, the HVR-H1 comprises an amino acid sequence with atleast about 90%, at least about 91%, at least about 92%, at least about93%, at least about 94%, at least about 95%, at least about 96%, atleast about 97%, at least about 98%, at least about 99%, or 100%identity to an amino acid sequence of SEQ ID NO: 1. In some embodiments,the HVR-H1 comprises an amino acid sequence containing substitutions(e.g., conservative substitutions, insertions, or deletions relative toan amino acid sequence of SEQ ID NO: 1), but retains the ability to bindto Sortilin. In certain embodiments, up to 1, up to 2, up to 3, up to 4,or up to 5 amino acids been substituted, inserted, and/or deleted in theHVR-H1 amino acid sequence of SEQ ID NO: 1. In some embodiments, theHVR-H2 comprises an amino acid sequence with at least about 90%, atleast about 91%, at least about 92%, at least about 93%, at least about94%, at least about 95%, at least about 96%, at least about 97%, atleast about 98%, at least about 99%, or 100% identity to an amino acidsequence selected from SEQ ID NOs: 2-3. In some embodiments, the HVR-H2comprises an amino acid sequence containing substitutions (e.g.,conservative substitutions, insertions, or deletions relative to anamino acid sequence selected from SEQ ID NOs: 2-3), but retains theability to bind to Sortilin. In certain embodiments, up to 1, up to 2,up to 3, up to 4, or up to 5 amino acids been substituted, inserted,and/or deleted in the HVR-H2 amino acid sequence selected from SEQ IDNOs: 2-3. In some embodiments, the HVR-H3 comprises an amino acidsequence with at least about 90%, at least about 91%, at least about92%, at least about 93%, at least about 94%, at least about 95%, atleast about 96%, at least about 97%, at least about 98%, at least about99%, or 100% identity to an amino acid sequence selected from SEQ IDNOs: 5-6. In some embodiments, the HVR-H3 comprises an amino acidsequence containing substitutions (e.g., conservative substitutions,insertions, or deletions relative to an amino acid sequence selectedfrom SEQ ID NOs: 5-6), but retains the ability to bind to Sortilin. Incertain embodiments, up to 1, up to 2, up to 3, up to 4, or up to 5amino acids been substituted, inserted, and/or deleted in the HVR-H3amino acid sequence selected from SEQ ID NOs: 5-6.

In some embodiments, the heavy chain variable region comprises an HVR-H1comprising a sequence of YSISSGYYWG (SEQ ID NO: 1), an HVR-H2 comprisinga sequence according to Formula I, and an HVR-H3 comprising a sequenceaccording to Formula II.

In some embodiments, the heavy chain variable region comprises an HVR-H1comprising a sequence of SEQ ID NO: 1, an HVR-H2 comprising a sequenceselected from SEQ ID NOs: 2-3, and an HVR-H3 comprising a sequenceselected from SEQ ID NOs: 5-6.

In some embodiments, the heavy chain variable region comprises theHVR-H1, HVR-H2, and HVR-H3 of antibody S-60-10, S-60-11, S-60-12,S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2[N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6[N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10[N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I],S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17[N33L], S-60-16; S-60-18, S-60-19, S-60-24, or any combination thereof(as shown in Tables 14-16).

In some embodiments, anti-Sortilin antibodies of the present disclosureinclude a heavy chain variable region, wherein the heavy chain variableregion comprises one or more of: (a) an HVR-H1 comprising an amino acidsequence with at least 85%, at least 86%, at least 87%, at least 88%, atleast 89%, at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to an HVR-H1 amino acid sequence of antibodyS-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)],S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R],S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q],S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12[N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A],S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, orS-60-24; (b) an HVR-H2 comprising an amino acid sequence with at least85%, at least 86%, at least 87%, at least 88%, at least 89%, at least90%, at least 91%, at least 92%, at least 93%, at least 94%, at least95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%identity to an HVR-H2 amino acid sequence of antibody S-60-10, S-60-11,S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T],S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D],S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y],S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13[N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M],S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24; and (c) anHVR-H3 comprising an amino acid sequence with at least 85%, at least86%, at least 87%, at least 88%, at least 89%, at least 90%, at least91%, at least 92%, at least 93%, at least 94%, at least 95%, at least96%, at least 97%, at least 98%, at least 99%, or 100% identity to anHVR-H3 amino acid sequence of antibody S-60-10, S-60-11, S-60-12,S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2[N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6[N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10[N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I],S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17[N33L], S-60-16; S-60-18, S-60-19, or S-60-24.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise an HVR-H1 comprising the amino acid sequence YSISSGYYWG (SEQ IDNO: 1), an HVR-H2 comprising the amino acid sequence TIYHSGSTYYNPSLKS(SEQ ID NO: 2), and an HVR-H3 comprising the amino acid sequenceARQGSIKQGYYGMDV (SEQ ID NO: 6).

B. Light chain HVRs

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a light chain variable region comprising one or more (e.g., oneor more, two or more, or all three) HVRs selected from HVR-L1, HVR-L2,and HVR-L3 (as shown in Tables 17-19). In some embodiments, the lightchain variable region comprises an HVR-L1, an HVR-L2, and an HVR-L3 (asshown in Tables 17-19).

In some embodiments, the HVR-L1 comprises a sequence according toFormula III: RSSQX₁LLX₂SX₃GYNYLD (SEQ ID NO: 28), wherein X₁ is S or G,X₂ is R or H, and X₃ is N, T, S, G, R, D, H, K, Q, Y, E, W, F, I, V, A,M, or L. In some embodiments, the HVR-L1 comprises a sequence selectedfrom SEQ ID NOs: 8-27. In some embodiments, the HVR-L1 comprises asequence of RSSQSLLRSNGYNYLD (SEQ ID NO:8), RSSQSLLRSTGYNYLD (SEQ IDNO:9), RSSQS LLRSSGYNYLD (SEQ ID NO:10), RSSQSLLRSGGYNYLD (SEQ IDNO:11), RSSQSLLRSRG YNYLD (SEQ ID NO:12), RSSQSLLRSDGYNYLD (SEQ IDNO:13), RSSQSLLRSHGYNYLD (SEQ ID NO:14), RSSQSLLRSKGYNYLD (SEQ IDNO:15), RSSQSLLRSQGYNYLD (SEQ ID NO:16), RSSQSLLRSYGYNYLD (SEQ IDNO:17), RSSQSLLRSEGYNYLD (SEQ ID NO:18), RSSQSLLRSWGYNYLD (SEQ IDNO:19), RSSQSLLRSFGYNYLD (SEQ ID NO:20), RSSQSL LRSIGYNYLD (SEQ IDNO:21), RSSQSLLRSVGYNYLD (SEQ ID NO:22), RSSQSLLRSAG YNYLD (SEQ IDNO:23), RSSQSLLRSMGYNYLD (SEQ ID NO:24), RSSQSLLRSLGYNYLD (SEQ IDNO:25), RSSQSLLHSNGYNYLD (SEQ ID NO:26), or RSSQGLLRSNGYNYLD (SEQ IDNO:27). In one specific embodiment, the HVR-L1 comprises a sequence ofRSSQSLLRSNGYNYLD (SEQ ID NO:8). In another specific embodiment, theHVR-L1 comprises a sequence of RSSQSLLRSTGYNYLD (SEQ ID NO:9) (as shownin Table 19).

In some embodiments, the HVR-L2 comprises a sequence according toFormula IV: LGSNRX1S (SEQ ID NO: 31), wherein X1 is A or V. In someembodiments, the HVR-L2 comprises a sequence selected from SEQ ID NOs:29-30.

In some embodiments, the HVR-L3 comprises a sequence according toFormula V: MQQQEX1PLT (SEQ ID NO: 34), wherein X1 is A or T. In someembodiments, the HVR-L3 comprises a sequence selected from SEQ ID NOs:32-33.

In some embodiments, the HVR-L1 comprises an amino acid sequence with atleast about 90%, at least about 91%, at least about 92%, at least about93%, at least about 94%, at least about 95%, at least about 96%, atleast about 97%, at least about 98%, at least about 99%, or 100%identity to an amino acid sequence selected from SEQ ID NOs: 8-27. Insome embodiments, the HVR-L1 comprises an amino acid sequence containingsubstitutions (e.g., conservative substitutions, insertions, ordeletions relative to an amino acid sequence selected from SEQ ID NOs:8-27), but retains the ability to bind to Sortilin. In certainembodiments, up to 1, up to 2, up to 3, up to 4, or up to 5 amino acidsbeen substituted, inserted, and/or deleted in the HVR-L1 amino acidsequence selected from SEQ ID NOs: 8-27. In some embodiments, the HVR-L2comprises an amino acid sequence with at least about 90%, at least about91%, at least about 92%, at least about 93%, at least about 94%, atleast about 95%, at least about 96%, at least about 97%, at least about98%, at least about 99%, or 100% identity to an amino acid sequenceselected from SEQ ID NOs: 29-30. In some embodiments, the HVR-L2comprises an amino acid sequence containing substitutions (e.g.,conservative substitutions, insertions, or deletions relative to anamino acid sequence selected from SEQ ID NOs: 29-30), but retains theability to bind to Sortilin. In certain embodiments, up to 1, up to 2,up to 3, up to 4, or up to 5 amino acids been substituted, inserted,and/or deleted in the HVR-L2 amino acid sequence selected from SEQ IDNOs: 29-30. In some embodiments, the HVR-L3 comprises an amino acidsequence with at least about 90%, at least about 91%, at least about92%, at least about 93%, at least about 94%, at least about 95%, atleast about 96%, at least about 97%, at least about 98%, at least about99%, or 100% identity to an amino acid sequence selected from SEQ IDNOs: 32-33. In some embodiments, the HVR-L3 comprises an amino acidsequence containing substitutions (e.g., conservative substitutions,insertions, or deletions relative to an amino acid sequence selectedfrom SEQ ID NOs: 32-33), but retains the ability to bind to Sortilin. Incertain embodiments, up to 1, up to 2, up to 3, up to 4, or up to 5amino acids been substituted, inserted, and/or deleted in the HVR-L3amino acid sequence selected from SEQ ID NOs: 32-33.

In some embodiments, the light chain variable region comprises an HVR-L1comprising a sequence according to Formula III, an HVR-L2 comprising asequence according to Formula IV, and an HVR-L3 comprising a sequenceaccording to Formula V. In some embodiments, the light chain variableregion comprises an HVR-L1 comprising a sequence selected from SEQ IDNOs: 8-27, an HVR-L2 comprising a sequence selected from SEQ ID NOs:29-30, and an HVR-L3 comprising a sequence selected from SEQ ID NOs:32-33.

In some embodiments, the light chain variable region comprises theHVR-L1, HVR-L2, and HVR-L3 of antibody S-60-10, S-60-11, S-60-12,S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2[N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6[N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10[N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I],S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17[N33L], S-60-16; S-60-18, S-60-19, S-60-24, or any combination thereof(as shown in Tables 17-19).

In some embodiments, anti-Sortilin antibodies of the present disclosureinclude a light chain variable region, wherein the light chain variableregion comprises one or more of: (a) an HVR-L1 comprising an amino acidsequence with at least 85%, at least 86%, at least 87%, at least 88%, atleast 89%, at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to an HVR-L1 amino acid sequence of antibodyS-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)],S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R],S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q],S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12[N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A],S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, orS-60-24; (b) an HVR-L2 comprising an amino acid sequence with at least85%, at least 86%, at least 87%, at least 88%, at least 89%, at least90%, at least 91%, at least 92%, at least 93%, at least 94%, at least95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%identity to an HVR-L2 amino acid sequence of antibody S-60-10, S-60-11,S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T],S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D],S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y],S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13[N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M],S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24; and (c) anHVR-L3 comprising an amino acid sequence with at least 85%, at least86%, at least 87%, at least 88%, at least 89%, at least 90%, at least91%, at least 92%, at least 93%, at least 94%, at least 95%, at least96%, at least 97%, at least 98%, at least 99%, or 100% identity to anHVR-L3 amino acid sequence of antibody S-60-10, S-60-11, S-60-12,S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2[N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6[N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10[N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I],S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17[N33L], S-60-16; S-60-18, S-60-19, or S-60-24.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise an HVR-L1 comprising the amino acid sequence RSSQSLLRSNGYNYLD(SEQ ID NO: 8), an HVR-L2 comprising the amino acid sequence LGSNRAS(SEQ ID NO: 29), and an HVR-L3 comprising the amino acid sequenceMQQQEAPLT (SEQ ID NO: 32).

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise an HVR-L1 comprising the amino acid sequence RSSQSLLRSTGYNYLD(SEQ ID NO: 9), an HVR-L2 comprising the amino acid sequence LGSNRAS(SEQ ID NO: 29), and an HVR-L3 comprising the amino acid sequenceMQQQEAPLT (SEQ ID NO: 32).

C. Heavy Chain HVRs and Light Chain HVRs

In some embodiments, anti-Sortilin antibodies of the present disclosureinclude a heavy chain variable region comprising one or more (e.g., oneor more, two or more, or all three) HVRs selected from HVR-H1, HVR-H2,and HVR-H3 (as shown in Tables 14-16), and a light chain variable regioncomprising one or more (e.g., one or more, two or more, or all three)HVRs selected from HVR-L1, HVR-L2, and HVR-L3 (as shown in Tables17-19). In some embodiments, the heavy chain variable region comprisesan HVR-H1, an HVR-H2, and an HVR-H3 (as shown in Tables 14-16), and thelight chain variable region comprises an HVR-L1, an HVR-L2, and anHVR-L3 (as shown in Tables 17-19).

In some embodiments, the heavy chain variable region comprises an HVR-H1comprising a sequence of YSISSGYYWG (SEQ ID NO: 1), an HVR-H2 comprisinga sequence according to Formula I, and an HVR-H3 comprising a sequenceaccording to Formula II, and the light chain variable region comprisesan HVR-L1 comprising a sequence according to Formula III, an HVR-L2comprising a sequence according to Formula IV, and an HVR-L3 comprisinga sequence according to Formula V. In some embodiments, the heavy chainvariable region comprises an HVR-H1 comprising a sequence of SEQ ID NO:1, an HVR-H2 comprising a sequence selected from SEQ ID NOs: 2-3, and anHVR-H3 comprising a sequence selected from SEQ ID NOs: 5-6, and thelight chain variable region comprises an HVR-L1 comprising a sequenceselected from SEQ ID NOs: 8-27, an HVR-L2 comprising a sequence selectedfrom SEQ ID NOs: 29-30, and an HVR-L3 comprising a sequence selectedfrom SEQ ID NOs: 32-33.

In some aspects, the heavy chain variable region comprises an HVR-H1comprising a sequence of SEQ ID NO: 1, an HVR-H2 comprising a sequenceselected from SEQ ID NOs: 2-3, and an HVR-H3 comprising a sequenceselected from SEQ ID NOs: 5-6, and the light chain variable regioncomprises an HVR-L1 comprising a sequence selected from SEQ ID NOs:8-27, an HVR-L2 comprising a sequence selected from SEQ ID NOs: 29-30,and an HVR-L3 comprising a sequence of SEQ ID NO: 32.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable region comprising the HVR-H1, HVR-H2,and HVR-H3 of antibody S-60-10, S-60-11, S-60-12, S-60-13, S-60-14,S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3[N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7[N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E],S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14[N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L],S-60-16; S-60-18, S-60-19, S-60-24, or any combination thereof (as shownin Tables 14-16); and a light chain variable region comprising theHVR-L1, HVR-L2, and HVR-L3 of antibody S-60-10, S-60-11, S-60-12,S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2[N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6[N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10[N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I],S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17[N33L], S-60-16; S-60-18, S-60-19, S-60-24, or any combination thereof(as shown in Tables 17-19).

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable region comprising an HVR-H1, HVR-H2, andHVR-H3 and a light chain variable region comprising an HVR-L1, HVR-L2,and HVR-L3, wherein the antibody comprises the HVR-H1, HVR-H2, HVR-H3,HVR-L1, HVR-L2, and HVR-L3 of antibody S-60-10, S-60-11, S-60-12,S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2[N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6[N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10[N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I],S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17[N33L], S-60-16; S-60-18, S-60-19, or S-60-24 (as shown in Tables14-19).

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable region and a light chain variableregion, wherein the heavy chain variable region comprises one or moreof: (a) an HVR-H1 comprising an amino acid sequence with at least 90%,at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, atleast 96%, at least 97%, at least 98%, at least 99%, or 100% identity toan HVR-H1 amino acid sequence of antibody S-60-10, S-60-11, S-60-12,S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2[N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6[N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10[N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I],S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17[N33L], S-60-16; S-60-18, S-60-19, or S-60-24; (b) an HVR-H2 comprisingan amino acid sequence with at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to an HVR-H2 amino acidsequence of antibody S-60-10, S-60-11, S-60-12, S-60-13, S-60-14,S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3[N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7[N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E],S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14[N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L],S-60-16; S-60-18, S-60-19, or S-60-24; and (c) an HVR-H3 comprising anamino acid sequence with at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to an HVR-H3 amino acidsequence of antibody S-60-10, S-60-11, S-60-12, S-60-13, S-60-14,S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3[N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7[N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E],S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14[N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L],S-60-16; S-60-18, S-60-19, or S-60-24; and wherein the light chainvariable region comprises one or more of: (a) an HVR-L1 comprising anamino acid sequence with at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to an HVR-L1 amino acidsequence of antibody S-60-10, S-60-11, S-60-12, S-60-13, S-60-14,S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3[N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7[N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E],S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14[N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L],S-60-16; S-60-18, S-60-19, or S-60-24; (b) an HVR-L2 comprising an aminoacid sequence with at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, at least 99%, or 100% identity to an HVR-L2 amino acid sequence ofantibody S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33(wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4[N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8[N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W],S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15[N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19,or S-60-24; and (c) an HVR-L3 comprising an amino acid sequence with atleast 90%, at least 91%, at least 92%, at least 93%, at least 94%, atleast 95%, at least 96%, at least 97%, at least 98%, at least 99%, or100% identity to an HVR-L3 amino acid sequence of antibody S-60-10,S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1[N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5[N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9[N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F],S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16[N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24.

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the HVR-H1 comprisingthe amino acid sequence YSISSGYYWG (SEQ ID NO: 1), the HVR-H2 comprisingthe amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), the HVR-H3comprising the amino acid sequence ARQGSIQQGYYGMDV (SEQ ID NO: 5); and alight chain variable region comprising the HVR-L1 comprising the aminoacid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), the HVR-L2 comprising theamino acid sequence LGSNRAS (SEQ ID NO: 29), and the HVR-L3 comprisingthe amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the HVR-H1 comprisingthe amino acid sequence YSISSGYYWG (SEQ ID NO: 1), the HVR-H2 comprisingthe amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), the HVR-H3comprising the amino acid sequence ARQGSIQQGYYGMDV (SEQ ID NO: 5); and alight chain variable region comprising the HVR-L1 comprising the aminoacid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), the HVR-L2 comprising theamino acid sequence LGSNRVS (SEQ ID NO: 30), and the HVR-L3 comprisingthe amino acid sequence MQQQETPLT (SEQ ID NO: 33).

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the HVR-H1 comprisingthe amino acid sequence YSISSGYYWG (SEQ ID NO: 1), the HVR-H2 comprisingthe amino acid sequence TIYHSGSTYYNPSLES (SEQ ID NO: 3), the HVR-H3comprising the amino acid sequence ARQGSIQQGYYGMDV (SEQ ID NO: 5); and alight chain variable region comprising the HVR-L1 comprising the aminoacid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), the HVR-L2 comprising theamino acid sequence LGSNRAS (SEQ ID NO: 29), and the HVR-L3 comprisingthe amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some aspects, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the HVR-H1 comprisingthe amino acid sequence YSISSGYYWG (SEQ ID NO: 1), the HVR-H2 comprisingthe amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), the HVR-H3comprising the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and alight chain variable region comprising the HVR-L1 comprising the aminoacid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), the HVR-L2 comprising theamino acid sequence LGSNRAS (SEQ ID NO: 29), and the HVR-L3 comprisingthe amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some aspects, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the HVR-H1 comprisingthe amino acid sequence YSISSGYYWG (SEQ ID NO: 1), the HVR-H2 comprisingthe amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), the HVR-H3comprising the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and alight chain variable region comprising the HVR-L1 comprising the aminoacid sequence RSSQSLLRSTGYNYLD (SEQ ID NO: 9), the HVR-L2 comprising theamino acid sequence LGSNRAS (SEQ ID NO: 29), and the HVR-L3 comprisingthe amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the HVR-H1 comprisingthe amino acid sequence YSISSGYYWG (SEQ ID NO: 1), the HVR-H2 comprisingthe amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), the HVR-H3comprising the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and alight chain variable region comprising the HVR-L1 comprising the aminoacid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), the HVR-L2 comprising theamino acid sequence LGSNRAS (SEQ ID NO: 29), and the HVR-L3 comprisingthe amino acid sequence MQQQETPLT (SEQ ID NO: 33).

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the HVR-H1 comprisingthe amino acid sequence YSISSGYYWG (SEQ ID NO: 1), the HVR-H2 comprisingthe amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), the HVR-H3comprising the amino acid sequence ARQGSIQQGYYGMDV (SEQ ID NO: 5); and alight chain variable region comprising the HVR-L1 comprising the aminoacid sequence RSSQSLLHSNGYNYLD (SEQ ID NO: 26), the HVR-L2 comprisingthe amino acid sequence LGSNRAS (SEQ ID NO: 29), and the HVR-L3comprising the amino acid sequence MQQQETPLT (SEQ ID NO: 33).

In some embodiments, an anti-Sortilin antibody of the present disclosurecomprises a heavy chain variable region comprising the HVR-H1 comprisingthe amino acid sequence YSISSGYYWG (SEQ ID NO: 1), the HVR-H2 comprisingthe amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), the HVR-H3comprising the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and alight chain variable region comprising the HVR-L1 comprising the aminoacid sequence RSSQGLLRSNGYNYLD (SEQ ID NO: 27), the HVR-L2 comprisingthe amino acid sequence LGSNRAS (SEQ ID NO: 29), and the HVR-L3comprising the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

D. Heavy Chain Variable Region

In some embodiments, anti-Sortilin antibodies of the present disclosureinclude a heavy chain variable region comprising an amino acid sequenceselected from SEQ ID NOs: 54-56. In some embodiments, the heavy chainvariable region comprises an amino acid sequence with at least about90%, at least about 91%, at least about 92%, at least about 93%, atleast about 94%, at least about 95%, at least about 96%, at least about97%, at least about 98%, at least about 99%, or 100% identity to anamino acid sequence selected from SEQ ID NOs: 54-56. In someembodiments, the heavy chain variable region comprises an amino acidsequence containing substitutions (e.g., conservative substitutions,insertions, or deletions relative to an amino acid sequence selectedfrom SEQ ID NOs: 54-56), but retains the ability to bind to Sortilin. Incertain embodiments, up to 1, up to 2, up to 3, up to 4, up to 5, up to6, up to 7, up to 8, up to 9, or up to 10 amino acids been substituted,inserted, and/or deleted in the heavy chain variable region amino acidsequence selected from SEQ ID NOs: 54-56.

In some embodiments, the heavy chain variable region comprises the aminoacid sequence of SEQ ID NO: 56.

In some embodiments, anti-Sortilin antibodies of the present disclosureinclude a heavy chain variable region of antibody S-60-10, S-60-11,S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T],S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D],S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y],S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13[N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M],S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24 (as shown inTable 30).

In some embodiments, anti-Sortilin antibodies of the present disclosureinclude a heavy chain variable region comprising an HVR-H1 comprisingthe amino acid sequence YSISSGYYWG (SEQ ID NO: 1), an HVR-H2 comprisingthe amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), and an HVR-H3comprising the amino acid sequence ARQGSIKQGYYGMDV (SEQ ID NO: 6).

E. Light Chain Variable Region

In some embodiments, anti-Sortilin antibodies of the present disclosureinclude a light chain variable region comprising an amino acid sequenceselected from SEQ ID NOs: 57-80. In some embodiments, the light chainvariable region comprises an amino acid sequence with at least about90%, at least about 91%, at least about 92%, at least about 93%, atleast about 94%, at least about 95%, at least about 96%, at least about97%, at least about 98%, at least about 99%, or 100% identity to anamino acid sequence selected from SEQ ID NOs: 57-80. In someembodiments, the light chain variable region comprises an amino acidsequence containing substitutions (e.g., conservative substitutions,insertions, or deletions relative to an amino acid sequence selectedfrom SEQ ID NOs: 57-80), but retains the ability to bind to Sortilin. Incertain embodiments, up to 1, up to 2, up to 3, up to 4, up to 5, up to6, up to 7, up to 8, up to 9, or up to 10 amino acids been substituted,inserted, and/or deleted in the light chain variable region amino acidsequence selected from SEQ ID NOs: 57-80.

In some embodiments, the light chain variable region includes the aminoacid sequence of SEQ ID NO: 57. In some embodiments, the light chainvariable region includes the amino acid sequence of SEQ ID NO: 60.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a light chain variable region of antibody S-60-10, S-60-11,S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T],S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D],S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y],S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13[N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M],S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24 (as shown inTable 31).

In some embodiments, anti-Sortilin antibodies of the present disclosureinclude a light chain variable region comprising an HVR-L1 comprisingthe amino acid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), an HVR-L2comprising the amino acid sequence LGSNRAS (SEQ ID NO: 29), and anHVR-L3 comprising the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

In some embodiments, anti-Sortilin antibodies of the present disclosureinclude a light chain variable region comprising an HVR-L1 comprisingthe amino acid sequence RSSQSLLRSTGYNYLD (SEQ ID NO: 9), an HVR-L2comprising the amino acid sequence LGSNRAS (SEQ ID NO: 29), and anHVR-L3 comprising the amino acid sequence MQQQEAPLT (SEQ ID NO: 32).

F. Heavy Chain Variable Region and Light Chain Variable Region

In some aspects, an anti-Sortilin antibody of the present disclosureincludes a heavy chain variable region comprising an amino acid sequenceselected from the group consisting of SEQ ID NOs: 54-56; and/or a lightchain variable region comprises an amino acid sequence selected from thegroup consisting of SEQ ID NOs: 57-80. In some embodiments, the heavychain variable region comprises an amino acid sequence with at leastabout 90%, at least about 91%, at least about 92%, at least about 93%,at least about 94%, at least about 95%, at least about 96%, at leastabout 97%, at least about 98%, at least about 99%, or 100% identity toan amino acid sequence selected from SEQ ID NOs: 54-56, and the lightchain variable region comprises an amino acid sequence with at leastabout 90%, at least about 91%, at least about 92%, at least about 93%,at least about 94%, at least about 95%, at least about 96%, at leastabout 97%, at least about 98%, at least about 99%, or 100% identity toan amino acid sequence selected from SEQ ID NOs: 57-80. In someembodiments, an anti-Sortilin antibody of the present disclosureincludes a heavy chain variable region comprising an amino acid sequencecontaining substitutions (e.g., conservative substitutions, insertions,or deletions relative to an amino acid sequence selected from SEQ IDNOs: 54-56), and a light chain variable region comprising an amino acidsequence containing substitutions (e.g., conservative substitutions,insertions, or deletions relative to an amino acid sequence selectedfrom SEQ ID NOs: 57-80), but retains the ability to bind to Sortilin. Incertain embodiments, up to 1, up to 2, up to 3, up to 4, up to 5, up to6, up to 7, up to 8, up to 9, or up to 10 amino acids been substituted,inserted, and/or deleted in the heavy chain variable region amino acidsequence selected from SEQ ID NOs: 54-56; and up to 1, up to 2, up to 3,up to 4, up to 5, up to 6, up to 7, up to 8, up to 9, or up to 10 aminoacids been substituted, inserted, and/or deleted in the light chainvariable region amino acid sequence selected from SEQ ID NOs: 57-80.

In some aspects, an anti-Sortilin antibody of the present disclosureincludes a heavy chain variable region comprising an amino acid sequenceselected from the group consisting of SEQ ID NOs: 54-56; and/or a lightchain variable region comprises an amino acid sequence selected from thegroup consisting of SEQ ID NOs: 57-58, 60-78, and 80.

In some embodiments, an anti-Sortilin antibody of the present disclosurebinds to a Sortilin protein, wherein the antibody includes a heavy chainvariable region comprising the amino acid sequence of SEQ ID NO: 54, anda light chain variable region comprising the amino acid sequence of SEQID NO: 57; a heavy chain variable region comprising the amino acidsequence of SEQ ID NO: 54, and a light chain variable region comprisingthe amino acid sequence of SEQ ID NO: 58; a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO: 54, and a light chainvariable region comprising the amino acid sequence of SEQ ID NO: 59; aheavy chain variable region comprising the amino acid sequence of SEQ IDNO: 55, and a light chain variable region comprising the amino acidsequence of SEQ ID NO: 57; a heavy chain variable region comprising theamino acid sequence of SEQ ID NO: 55, and a light chain variable regioncomprising the amino acid sequence of SEQ ID NO: 58; a heavy chainvariable region comprising the amino acid sequence of SEQ ID NO: 56, anda light chain variable region comprising the amino acid sequence of SEQID NO: 57; a heavy chain variable region comprising the amino acidsequence of SEQ ID NO: 56, and a light chain variable region comprisingthe amino acid sequence of SEQ ID NO: 77; a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO: 56, and a light chainvariable region comprising the amino acid sequence of SEQ ID NO: 78; aheavy chain variable region comprising the amino acid sequence of SEQ IDNO: 54, and a light chain variable region comprising the amino acidsequence of SEQ ID NO: 79; or a heavy chain variable region comprisingthe amino acid sequence of SEQ ID NO: 56, and a light chain variableregion comprising the amino acid sequence of SEQ ID NO: 80.

In one aspect, an anti-Sortilin antibody of the present disclosureincludes a heavy chain variable region having the amino acid sequence ofSEQ ID NO: 56, and a light chain variable region having the amino acidsequence of SEQ ID NO: 57.

In one aspect, an anti-Sortilin antibody of the present disclosureincludes a heavy chain variable region having the amino acid sequence ofSEQ ID NO: 56, and a light chain variable region having the amino acidsequence of SEQ ID NO: 60.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable region of antibody S-60-10, S-60-11,S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T],S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D],S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y],S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13[N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M],S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24 (as shown inTable 30), and a light chain variable region of antibody S-60-10,S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1[N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5[N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9[N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F],S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16[N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24 (asshown in Table 31).

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable region comprising the amino acidsequence of SEQ ID NO: 56, and a light chain variable region comprisingan amino acid sequence selected from SEQ ID NOs: 56 and 60. In someembodiments, the antibody comprises a heavy chain variable region ofS-60-15 [N33 (wt)] (as shown in Table 30), and a light chain variableregion of antibody S-60-15 [N33 (wt)] (as shown in Table 31). In someembodiments, the antibody comprises a heavy chain variable region ofS-60-15.1 [N33T] (as shown in Table 30), and a light chain variableregion of antibody S-60-15.1 [N33T] (as shown in Table 31).

Exemplary Anti-Sortilin Antibodies

In some embodiments, the anti-Sortilin antibody is an anti-Sortilinmonoclonal antibody comprising the heavy chain variable region and thelight chain variable region of an antibody selected from S-60-10,S-60-11, S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1[N33T], S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5[N33D], S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9[N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F],S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16[N33M], S-60-15.17 [N33L], S-60-16; S-60-18, S-60-19, or S-60-24. Insome embodiments, the anti-Sortilin antibody is an anti-Sortilinmonoclonal antibody comprising the heavy chain and the light chain of anantibody selected from S-60-10, S-60-11, S-60-12, S-60-13, S-60-14,S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3[N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7[N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E],S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14[N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L],S-60-16; S-60-18, S-60-19, or S-60-24.

(1) S-60-10

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable domain and a light chain variabledomain, wherein the heavy chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a heavy chain variable domain amino acidsequence of antibody S-60-10 or to the amino acid sequence of SEQ ID NO:54; and/or the light chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-10 or to the amino acid sequence of SEQ ID NO:57. In some embodiments, anti-Sortilin antibodies of the presentdisclosure comprise a heavy chain variable domain comprising an aminoacid sequence with at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, at least 99%, or 100% identity to a heavy chain variable domainamino acid sequence of antibody S-60-10 or to the amino acid sequence ofSEQ ID NO: 54, wherein the heavy chain variable domain comprises theHVR-H1, HVR-H2, and HVR-H3 amino acid sequences of antibody S-60-10. Insome embodiments, anti-Sortilin antibodies of the present disclosurecomprise a light chain variable domain comprising an amino acid sequencewith at least 90%, at least 91%, at least 92%, at least 93%, at least94%, at least 95%, at least 96%, at least 97%, at least 98%, at least99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-10 or to the amino acid sequence of SEQ ID NO:57, wherein the light chain variable domain comprises the HVR-L1,HVR-L2, and HVR-L3 amino acid sequences of antibody S-60-10. In someembodiments, the anti-Sortilin antibody comprises a heavy chain variabledomain (VH) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a heavy chain variabledomain amino acid sequence of antibody S-60-10 or to the amino acidsequence of SEQ ID NO: 54 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the heavy chain variable domain amino acid sequence of antibodyS-60-10 or the amino acid sequence of SEQ ID NO: 54. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the heavy chain variable domain amino acidsequence of antibody S-60-10 or the amino acid sequence of SEQ ID NO:54. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VHsequence of antibody S-60-10 or of SEQ ID NO: 54, includingpost-translational modifications of that sequence. In a particularembodiment, the VH comprises one, two or three HVRs selected from: (a)the HVR-H1 amino acid sequence of antibody S-60-10, (b) the HVR-H2 aminoacid sequence of antibody S-60-10, and (c) the HVR-H3 amino acidsequence of antibody S-60-10. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a light chain variabledomain (VL) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a light chain variabledomain amino acid sequence of antibody S-60-10 or to the amino acidsequence of SEQ ID NO: 57 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the light chain variable domain amino acid sequence of antibodyS-60-10 or the amino acid sequence of SEQ ID NO: 57. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the light chain variable domain amino acidsequence of antibody S-60-10 or the amino acid sequence of SEQ ID NO:57. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VLsequence of antibody S-60-10 or of SEQ ID NO: 57, includingpost-translational modifications of that sequence. In a particularembodiment, the VL comprises one, two or three HVRs selected from: (a)the HVR-L1 amino acid sequence of antibody S-60-10, (b) the HVR-L2 aminoacid sequence of antibody S-60-10, and (c) the HVR-L3 amino acidsequence of antibody S-60-10.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain comprising the amino acid sequence of SEQ ID NO:105 or SEQ ID NO: 106. In some embodiments, anti-Sortilin antibodies ofthe present disclosure comprise a light chain comprising the amino acidsequence of SEQ ID NO: 139. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a heavy chain comprisingthe amino acid sequence of SEQ ID NO: 105 or SEQ ID NO: 106 and a lightchain comprising the amino acid sequence of SEQ ID NO: 139.

(2) S-60-11

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable domain and a light chain variabledomain, wherein the heavy chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a heavy chain variable domain amino acidsequence of antibody S-60-11 or to the amino acid sequence of SEQ ID NO:54; and/or the light chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-11 or to the amino acid sequence of SEQ ID NO:58. In some embodiments, anti-Sortilin antibodies of the presentdisclosure comprise a heavy chain variable domain comprising an aminoacid sequence with at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, at least 99%, or 100% identity to a heavy chain variable domainamino acid sequence of antibody S-60-11 or to the amino acid sequence ofSEQ ID NO: 54, wherein the heavy chain variable domain comprises theHVR-H1, HVR-H2, and HVR-H3 amino acid sequences of antibody S-60-11. Insome embodiments, anti-Sortilin antibodies of the present disclosurecomprise a light chain variable domain comprising an amino acid sequencewith at least 90%, at least 91%, at least 92%, at least 93%, at least94%, at least 95%, at least 96%, at least 97%, at least 98%, at least99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-11 or to the amino acid sequence of SEQ ID NO:58, wherein the light chain variable domain comprises the HVR-L1,HVR-L2, and HVR-L3 amino acid sequences of antibody S-60-11. In someembodiments, the anti-Sortilin antibody comprises a heavy chain variabledomain (VH) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a heavy chain variabledomain amino acid sequence of antibody S-60-11 or to the amino acidsequence of SEQ ID NO: 54 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the heavy chain variable domain amino acid sequence of antibodyS-60-11 or the amino acid sequence of SEQ ID NO: 54. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the heavy chain variable domain amino acidsequence of antibody S-60-11 or the amino acid sequence of SEQ ID NO:54. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VHsequence of antibody S-60-11 or of SEQ ID NO: 54, includingpost-translational modifications of that sequence. In a particularembodiment, the VH comprises one, two or three HVRs selected from: (a)the HVR-H1 amino acid sequence of antibody S-60-11, (b) the HVR-H2 aminoacid sequence of antibody S-60-11, and (c) the HVR-H3 amino acidsequence of antibody S-60-11. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a light chain variabledomain (VL) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a light chain variabledomain amino acid sequence of antibody S-60-11 or to the amino acidsequence of SEQ ID NO: 58 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the light chain variable domain amino acid sequence of antibodyS-60-11 or the amino acid sequence of SEQ ID NO: 58. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the light chain variable domain amino acidsequence of antibody S-60-11 or the amino acid sequence of SEQ ID NO:58. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VLsequence of antibody S-60-11 or of SEQ ID NO: 58, includingpost-translational modifications of that sequence. In a particularembodiment, the VL comprises one, two or three HVRs selected from: (a)the HVR-L1 amino acid sequence of antibody S-60-11, (b) the HVR-L2 aminoacid sequence of antibody S-60-11, and (c) the HVR-L3 amino acidsequence of antibody S-60-11.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain comprising the amino acid sequence of SEQ ID NO:105 or SEQ ID NO: 106. In some embodiments, anti-Sortilin antibodies ofthe present disclosure comprise a light chain comprising the amino acidsequence of SEQ ID NO: 140. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a heavy chain comprisingthe amino acid sequence of SEQ ID NO: 105 or SEQ ID NO: 106 and a lightchain comprising the amino acid sequence of SEQ ID NO: 140.

(3) S-60-12

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable domain and a light chain variabledomain, wherein the heavy chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a heavy chain variable domain amino acidsequence of antibody S-60-12 or to the amino acid sequence of SEQ ID NO:54; and/or the light chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-12 or to the amino acid sequence of SEQ ID NO:59. In some embodiments, anti-Sortilin antibodies of the presentdisclosure comprise a heavy chain variable domain comprising an aminoacid sequence with at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, at least 99%, or 100% identity to a heavy chain variable domainamino acid sequence of antibody S-60-12 or to the amino acid sequence ofSEQ ID NO: 54, wherein the heavy chain variable domain comprises theHVR-H1, HVR-H2, and HVR-H3 amino acid sequences of antibody S-60-12. Insome embodiments, anti-Sortilin antibodies of the present disclosurecomprise a light chain variable domain comprising an amino acid sequencewith at least 90%, at least 91%, at least 92%, at least 93%, at least94%, at least 95%, at least 96%, at least 97%, at least 98%, at least99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-12 or to the amino acid sequence of SEQ ID NO:59, wherein the light chain variable domain comprises the HVR-L1,HVR-L2, and HVR-L3 amino acid sequences of antibody S-60-12. In someembodiments, the anti-Sortilin antibody comprises a heavy chain variabledomain (VH) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a heavy chain variabledomain amino acid sequence of antibody S-60-12 or to the amino acidsequence of SEQ ID NO: 54 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the heavy chain variable domain amino acid sequence of antibodyS-60-12 or the amino acid sequence of SEQ ID NO: 54. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the heavy chain variable domain amino acidsequence of antibody S-60-12 or the amino acid sequence of SEQ ID NO:54. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VHsequence of antibody S-60-12 or of SEQ ID NO: 54, includingpost-translational modifications of that sequence. In a particularembodiment, the VH comprises one, two or three HVRs selected from: (a)the HVR-H1 amino acid sequence of antibody S-60-12, (b) the HVR-H2 aminoacid sequence of antibody S-60-12, and (c) the HVR-H3 amino acidsequence of antibody S-60-12. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a light chain variabledomain (VL) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a light chain variabledomain amino acid sequence of antibody S-60-12 or to the amino acidsequence of SEQ ID NO: 59 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the light chain variable domain amino acid sequence of antibodyS-60-12 or the amino acid sequence of SEQ ID NO: 59. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the light chain variable domain amino acidsequence of antibody S-60-12 or the amino acid sequence of SEQ ID NO:59. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VLsequence of antibody S-60-12 or of SEQ ID NO: 59, includingpost-translational modifications of that sequence. In a particularembodiment, the VL comprises one, two or three HVRs selected from: (a)the HVR-L1 amino acid sequence of antibody S-60-12, (b) the HVR-L2 aminoacid sequence of antibody S-60-12, and (c) the HVR-L3 amino acidsequence of antibody S-60-12.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain comprising the amino acid sequence of SEQ ID NO:105 or SEQ ID NO: 106. In some embodiments, anti-Sortilin antibodies ofthe present disclosure comprise a light chain comprising the amino acidsequence of SEQ ID NO: 141. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a heavy chain comprisingthe amino acid sequence of SEQ ID NO: 105 or SEQ ID NO: 106 and a lightchain comprising the amino acid sequence of SEQ ID NO: 141.

(4) S-60-13

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable domain and a light chain variabledomain, wherein the heavy chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a heavy chain variable domain amino acidsequence of antibody S-60-13 or to the amino acid sequence of SEQ ID NO:55; and/or the light chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-13 or to the amino acid sequence of SEQ ID NO:57. In some embodiments, anti-Sortilin antibodies of the presentdisclosure comprise a heavy chain variable domain comprising an aminoacid sequence with at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, at least 99%, or 100% identity to a heavy chain variable domainamino acid sequence of antibody S-60-13 or to the amino acid sequence ofSEQ ID NO: 55, wherein the heavy chain variable domain comprises theHVR-H1, HVR-H2, and HVR-H3 amino acid sequences of antibody S-60-13. Insome embodiments, anti-Sortilin antibodies of the present disclosurecomprise a light chain variable domain comprising an amino acid sequencewith at least 90%, at least 91%, at least 92%, at least 93%, at least94%, at least 95%, at least 96%, at least 97%, at least 98%, at least99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-13 or to the amino acid sequence of SEQ ID NO:57, wherein the light chain variable domain comprises the HVR-L1,HVR-L2, and HVR-L3 amino acid sequences of antibody S-60-13. In someembodiments, the anti-Sortilin antibody comprises a heavy chain variabledomain (VH) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a heavy chain variabledomain amino acid sequence of antibody S-60-13 or to the amino acidsequence of SEQ ID NO: 55 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the heavy chain variable domain amino acid sequence of antibodyS-60-13 or the amino acid sequence of SEQ ID NO: 55. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the heavy chain variable domain amino acidsequence of antibody S-60-13 or the amino acid sequence of SEQ ID NO:55. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VHsequence of antibody S-60-13 or of SEQ ID NO: 55, includingpost-translational modifications of that sequence. In a particularembodiment, the VH comprises one, two or three HVRs selected from: (a)the HVR-H1 amino acid sequence of antibody S-60-13, (b) the HVR-H2 aminoacid sequence of antibody S-60-13, and (c) the HVR-H3 amino acidsequence of antibody S-60-13. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a light chain variabledomain (VL) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a light chain variabledomain amino acid sequence of antibody S-60-13 or to the amino acidsequence of SEQ ID NO: 57 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the light chain variable domain amino acid sequence of antibodyS-60-13 or the amino acid sequence of SEQ ID NO: 57. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the light chain variable domain amino acidsequence of antibody S-60-13 or the amino acid sequence of SEQ ID NO:57. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VLsequence of antibody S-60-13 or of SEQ ID NO: 57, includingpost-translational modifications of that sequence. In a particularembodiment, the VL comprises one, two or three HVRs selected from: (a)the HVR-L1 amino acid sequence of antibody S-60-13, (b) the HVR-L2 aminoacid sequence of antibody S-60-13, and (c) the HVR-L3 amino acidsequence of antibody S-60-13.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain comprising the amino acid sequence of SEQ ID NO:135 or SEQ ID NO: 136. In some embodiments, anti-Sortilin antibodies ofthe present disclosure comprise a light chain comprising the amino acidsequence of SEQ ID NO: 139. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a heavy chain comprisingthe amino acid sequence of SEQ ID NO: 135 or SEQ ID NO: 136 and a lightchain comprising the amino acid sequence of SEQ ID NO: 139.

(5) S-60-14

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable domain and a light chain variabledomain, wherein the heavy chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a heavy chain variable domain amino acidsequence of antibody S-60-14 or to the amino acid sequence of SEQ ID NO:55; and/or the light chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-14 or to the amino acid sequence of SEQ ID NO:58. In some embodiments, anti-Sortilin antibodies of the presentdisclosure comprise a heavy chain variable domain comprising an aminoacid sequence with at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, at least 99%, or 100% identity to a heavy chain variable domainamino acid sequence of antibody S-60-14 or to the amino acid sequence ofSEQ ID NO: 55, wherein the heavy chain variable domain comprises theHVR-H1, HVR-H2, and HVR-H3 amino acid sequences of antibody S-60-14. Insome embodiments, anti-Sortilin antibodies of the present disclosurecomprise a light chain variable domain comprising an amino acid sequencewith at least 90%, at least 91%, at least 92%, at least 93%, at least94%, at least 95%, at least 96%, at least 97%, at least 98%, at least99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-14 or to the amino acid sequence of SEQ ID NO:58, wherein the light chain variable domain comprises the HVR-L1,HVR-L2, and HVR-L3 amino acid sequences of antibody S-60-14. In someembodiments, the anti-Sortilin antibody comprises a heavy chain variabledomain (VH) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a heavy chain variabledomain amino acid sequence of antibody S-60-14 or to the amino acidsequence of SEQ ID NO: 55 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the heavy chain variable domain amino acid sequence of antibodyS-60-14 or the amino acid sequence of SEQ ID NO: 55. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the heavy chain variable domain amino acidsequence of antibody S-60-14 or the amino acid sequence of SEQ ID NO:55. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VHsequence of antibody S-60-14 or of SEQ ID NO: 55, includingpost-translational modifications of that sequence. In a particularembodiment, the VH comprises one, two or three HVRs selected from: (a)the HVR-H1 amino acid sequence of antibody S-60-14, (b) the HVR-H2 aminoacid sequence of antibody S-60-14, and (c) the HVR-H3 amino acidsequence of antibody S-60-14. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a light chain variabledomain (VL) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a light chain variabledomain amino acid sequence of antibody S-60-14 or to the amino acidsequence of SEQ ID NO: 58 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the light chain variable domain amino acid sequence of antibodyS-60-14 or the amino acid sequence of SEQ ID NO: 58. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the light chain variable domain amino acidsequence of antibody S-60-14 or the amino acid sequence of SEQ ID NO:58. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VLsequence of antibody S-60-14 or of SEQ ID NO: 58, includingpost-translational modifications of that sequence. In a particularembodiment, the VL comprises one, two or three HVRs selected from: (a)the HVR-L1 amino acid sequence of antibody S-60-14, (b) the HVR-L2 aminoacid sequence of antibody S-60-14, and (c) the HVR-L3 amino acidsequence of antibody S-60-14.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain comprising the amino acid sequence of SEQ ID NO:135 or SEQ ID NO: 136. In some embodiments, anti-Sortilin antibodies ofthe present disclosure comprise a light chain comprising the amino acidsequence of SEQ ID NO: 140. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a heavy chain comprisingthe amino acid sequence of SEQ ID NO: 135 or SEQ ID NO: 136 and a lightchain comprising the amino acid sequence of SEQ ID NO: 140.

(6) S-60-15

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable domain and a light chain variabledomain, wherein the heavy chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a heavy chain variable domain amino acidsequence of antibody S-60-15 or to the amino acid sequence of SEQ ID NO:56; and/or the light chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-15 or to the amino acid sequence of SEQ ID NO:57. In some embodiments, anti-Sortilin antibodies of the presentdisclosure comprise a heavy chain variable domain comprising an aminoacid sequence with at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, at least 99%, or 100% identity to a heavy chain variable domainamino acid sequence of antibody S-60-15 or to the amino acid sequence ofSEQ ID NO: 56, wherein the heavy chain variable domain comprises theHVR-H1, HVR-H2, and HVR-H3 amino acid sequences of antibody S-60-15. Insome embodiments, anti-Sortilin antibodies of the present disclosurecomprise a light chain variable domain comprising an amino acid sequencewith at least 90%, at least 91%, at least 92%, at least 93%, at least94%, at least 95%, at least 96%, at least 97%, at least 98%, at least99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-15 or to the amino acid sequence of SEQ ID NO:57, wherein the light chain variable domain comprises the HVR-L1,HVR-L2, and HVR-L3 amino acid sequences of antibody S-60-15. In someembodiments, the anti-Sortilin antibody comprises a heavy chain variabledomain (VH) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a heavy chain variabledomain amino acid sequence of antibody S-60-15 or to the amino acidsequence of SEQ ID NO: 56 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the heavy chain variable domain amino acid sequence of antibodyS-60-15 or the amino acid sequence of SEQ ID NO: 56. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the heavy chain variable domain amino acidsequence of antibody S-60-15 or the amino acid sequence of SEQ ID NO:56. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VHsequence of antibody S-60-15 or of SEQ ID NO: 56, includingpost-translational modifications of that sequence. In a particularembodiment, the VH comprises one, two or three HVRs selected from: (a)the HVR-H1 amino acid sequence of antibody S-60-15, (b) the HVR-H2 aminoacid sequence of antibody S-60-15, and (c) the HVR-H3 amino acidsequence of antibody S-60-15. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a light chain variabledomain (VL) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a light chain variabledomain amino acid sequence of antibody S-60-15 or to the amino acidsequence of SEQ ID NO: 57 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the light chain variable domain amino acid sequence of antibodyS-60-15 or the amino acid sequence of SEQ ID NO: 57. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the light chain variable domain amino acidsequence of antibody S-60-15 or the amino acid sequence of SEQ ID NO:57. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VLsequence of antibody S-60-15 or of SEQ ID NO: 57, includingpost-translational modifications of that sequence. In a particularembodiment, the VL comprises one, two or three HVRs selected from: (a)the HVR-L1 amino acid sequence of antibody S-60-15, (b) the HVR-L2 aminoacid sequence of antibody S-60-15, and (c) the HVR-L3 amino acidsequence of antibody S-60-15.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain comprising the amino acid sequence of SEQ ID NO:137 or SEQ ID NO: 138. In some embodiments, anti-Sortilin antibodies ofthe present disclosure comprise a light chain comprising the amino acidsequence of SEQ ID NO: 139. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a heavy chain comprisingthe amino acid sequence of SEQ ID NO: 137 or SEQ ID NO: 138 and a lightchain comprising the amino acid sequence of SEQ ID NO: 139.

(7) S-60-15.1

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable domain and a light chain variabledomain, wherein the heavy chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a heavy chain variable domain amino acidsequence of antibody S-60-15.1 or to the amino acid sequence of SEQ IDNO: 56; and/or the light chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-15.1 or to the amino acid sequence of SEQ IDNO: 60. In some embodiments, anti-Sortilin antibodies of the presentdisclosure comprise a heavy chain variable domain comprising an aminoacid sequence with at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, at least 99%, or 100% identity to a heavy chain variable domainamino acid sequence of antibody S-60-15.1 or to the amino acid sequenceof SEQ ID NO: 56, wherein the heavy chain variable domain comprises theHVR-H1, HVR-H2, and HVR-H3 amino acid sequences of antibody S-60-15.1.In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a light chain variable domain comprising an amino acid sequencewith at least 90%, at least 91%, at least 92%, at least 93%, at least94%, at least 95%, at least 96%, at least 97%, at least 98%, at least99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-15.1 or to the amino acid sequence of SEQ IDNO: 60, wherein the light chain variable domain comprises the HVR-L1,HVR-L2, and HVR-L3 amino acid sequences of antibody S-60-15.1. In someembodiments, the anti-Sortilin antibody comprises a heavy chain variabledomain (VH) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a heavy chain variabledomain amino acid sequence of antibody S-60-15.1 or to the amino acidsequence of SEQ ID NO: 56 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the heavy chain variable domain amino acid sequence of antibodyS-60-15.1 or the amino acid sequence of SEQ ID NO: 56. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the heavy chain variable domain amino acidsequence of antibody S-60-15.1 or the amino acid sequence of SEQ ID NO:56. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VHsequence of antibody S-60-15.1 or of SEQ ID NO: 56, includingpost-translational modifications of that sequence. In a particularembodiment, the VH comprises one, two or three HVRs selected from: (a)the HVR-H1 amino acid sequence of antibody S-60-15.1, (b) the HVR-H2amino acid sequence of antibody S-60-15.1, and (c) the HVR-H3 amino acidsequence of antibody S-60-15.1. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a light chain variabledomain (VL) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a light chain variabledomain amino acid sequence of antibody S-60-15.1 or to the amino acidsequence of SEQ ID NO: 60 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the light chain variable domain amino acid sequence of antibodyS-60-15.1 or the amino acid sequence of SEQ ID NO: 60. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the light chain variable domain amino acidsequence of antibody S-60-15.1 or the amino acid sequence of SEQ ID NO:60. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VLsequence of antibody S-60-15.1 or of SEQ ID NO: 60, includingpost-translational modifications of that sequence. In a particularembodiment, the VL comprises one, two or three HVRs selected from: (a)the HVR-L1 amino acid sequence of antibody S-60-15.1, (b) the HVR-L2amino acid sequence of antibody S-60-15.1, and (c) the HVR-L3 amino acidsequence of antibody S-60-15.1.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain comprising the amino acid sequence of SEQ ID NO:137 or SEQ ID NO: 138. In some embodiments, anti-Sortilin antibodies ofthe present disclosure comprise a light chain comprising the amino acidsequence of SEQ ID NO: 142. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a heavy chain comprisingthe amino acid sequence of SEQ ID NO: 137 or SEQ ID NO: 138 and a lightchain comprising the amino acid sequence of SEQ ID NO: 142.

(8) S-60-16

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable domain and a light chain variabledomain, wherein the heavy chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a heavy chain variable domain amino acidsequence of antibody S-60-16 or to the amino acid sequence of SEQ ID NO:56; and/or the light chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-16 or to the amino acid sequence of SEQ ID NO:77. In some embodiments, anti-Sortilin antibodies of the presentdisclosure comprise a heavy chain variable domain comprising an aminoacid sequence with at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, at least 99%, or 100% identity to a heavy chain variable domainamino acid sequence of antibody S-60-16 or to the amino acid sequence ofSEQ ID NO: 56, wherein the heavy chain variable domain comprises theHVR-H1, HVR-H2, and HVR-H3 amino acid sequences of antibody S-60-16. Insome embodiments, anti-Sortilin antibodies of the present disclosurecomprise a light chain variable domain comprising an amino acid sequencewith at least 90%, at least 91%, at least 92%, at least 93%, at least94%, at least 95%, at least 96%, at least 97%, at least 98%, at least99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-16 or to the amino acid sequence of SEQ ID NO:77, wherein the light chain variable domain comprises the HVR-L1,HVR-L2, and HVR-L3 amino acid sequences of antibody S-60-16. In someembodiments, the anti-Sortilin antibody comprises a heavy chain variabledomain (VH) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a heavy chain variabledomain amino acid sequence of antibody S-60-16 or to the amino acidsequence of SEQ ID NO: 56 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the heavy chain variable domain amino acid sequence of antibodyS-60-16 or the amino acid sequence of SEQ ID NO: 56. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the heavy chain variable domain amino acidsequence of antibody S-60-16 or the amino acid sequence of SEQ ID NO:56. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VHsequence of antibody S-60-16 or of SEQ ID NO: 56, includingpost-translational modifications of that sequence. In a particularembodiment, the VH comprises one, two or three HVRs selected from: (a)the HVR-H1 amino acid sequence of antibody S-60-16, (b) the HVR-H2 aminoacid sequence of antibody S-60-16, and (c) the HVR-H3 amino acidsequence of antibody S-60-16. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a light chain variabledomain (VL) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a light chain variabledomain amino acid sequence of antibody S-60-16 or to the amino acidsequence of SEQ ID NO: 77 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the light chain variable domain amino acid sequence of antibodyS-60-16 or the amino acid sequence of SEQ ID NO: 77. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the light chain variable domain amino acidsequence of antibody S-60-16 or the amino acid sequence of SEQ ID NO:77. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VLsequence of antibody S-60-16 or of SEQ ID NO: 77, includingpost-translational modifications of that sequence. In a particularembodiment, the VL comprises one, two or three HVRs selected from: (a)the HVR-L1 amino acid sequence of antibody S-60-16, (b) the HVR-L2 aminoacid sequence of antibody S-60-16, and (c) the HVR-L3 amino acidsequence of antibody S-60-16.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain comprising the amino acid sequence of SEQ ID NO:137 or SEQ ID NO: 138. In some embodiments, anti-Sortilin antibodies ofthe present disclosure comprise a light chain comprising the amino acidsequence of SEQ ID NO: 131. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a heavy chain comprisingthe amino acid sequence of SEQ ID NO: 137 or SEQ ID NO: 138 and a lightchain comprising the amino acid sequence of SEQ ID NO: 131.

(9) S-60-18

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable domain and a light chain variabledomain, wherein the heavy chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a heavy chain variable domain amino acidsequence of antibody S-60-18 or to the amino acid sequence of SEQ ID NO:56; and/or the light chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-18 or to the amino acid sequence of SEQ ID NO:78. In some embodiments, anti-Sortilin antibodies of the presentdisclosure comprise a heavy chain variable domain comprising an aminoacid sequence with at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, at least 99%, or 100% identity to a heavy chain variable domainamino acid sequence of antibody S-60-18 or to the amino acid sequence ofSEQ ID NO: 56, wherein the heavy chain variable domain comprises theHVR-H1, HVR-H2, and HVR-H3 amino acid sequences of antibody S-60-18. Insome embodiments, anti-Sortilin antibodies of the present disclosurecomprise a light chain variable domain comprising an amino acid sequencewith at least 90%, at least 91%, at least 92%, at least 93%, at least94%, at least 95%, at least 96%, at least 97%, at least 98%, at least99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-18 or to the amino acid sequence of SEQ ID NO:78, wherein the light chain variable domain comprises the HVR-L1,HVR-L2, and HVR-L3 amino acid sequences of antibody S-60-18. In someembodiments, the anti-Sortilin antibody comprises a heavy chain variabledomain (VH) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a heavy chain variabledomain amino acid sequence of antibody S-60-18 or to the amino acidsequence of SEQ ID NO: 56 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the heavy chain variable domain amino acid sequence of antibodyS-60-18 or the amino acid sequence of SEQ ID NO: 56. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the heavy chain variable domain amino acidsequence of antibody S-60-18 or the amino acid sequence of SEQ ID NO:56. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VHsequence of antibody S-60-18 or of SEQ ID NO: 56, includingpost-translational modifications of that sequence. In a particularembodiment, the VH comprises one, two or three HVRs selected from: (a)the HVR-H1 amino acid sequence of antibody S-60-18, (b) the HVR-H2 aminoacid sequence of antibody S-60-18, and (c) the HVR-H3 amino acidsequence of antibody S-60-18. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a light chain variabledomain (VL) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a light chain variabledomain amino acid sequence of antibody S-60-18 or to the amino acidsequence of SEQ ID NO: 78 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the light chain variable domain amino acid sequence of antibodyS-60-18 or the amino acid sequence of SEQ ID NO: 78. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the light chain variable domain amino acidsequence of antibody S-60-18 or the amino acid sequence of SEQ ID NO:78. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VLsequence of antibody S-60-18 or of SEQ ID NO: 78, includingpost-translational modifications of that sequence. In a particularembodiment, the VL comprises one, two or three HVRs selected from: (a)the HVR-L1 amino acid sequence of antibody S-60-18, (b) the HVR-L2 aminoacid sequence of antibody S-60-18, and (c) the HVR-L3 amino acidsequence of antibody S-60-18.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain comprising the amino acid sequence of SEQ ID NO:137 or SEQ ID NO: 138. In some embodiments, anti-Sortilin antibodies ofthe present disclosure comprise a light chain comprising the amino acidsequence of SEQ ID NO: 132. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a heavy chain comprisingthe amino acid sequence of SEQ ID NO: 137 or SEQ ID NO: 138 and a lightchain comprising the amino acid sequence of SEQ ID NO: 132.

(10) S-60-19

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable domain and a light chain variabledomain, wherein the heavy chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a heavy chain variable domain amino acidsequence of antibody S-60-19 or to the amino acid sequence of SEQ ID NO:54; and/or the light chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-19 or to the amino acid sequence of SEQ ID NO:79. In some embodiments, anti-Sortilin antibodies of the presentdisclosure comprise a heavy chain variable domain comprising an aminoacid sequence with at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, at least 99%, or 100% identity to a heavy chain variable domainamino acid sequence of antibody S-60-19 or to the amino acid sequence ofSEQ ID NO: 54, wherein the heavy chain variable domain comprises theHVR-H1, HVR-H2, and HVR-H3 amino acid sequences of antibody S-60-19. Insome embodiments, anti-Sortilin antibodies of the present disclosurecomprise a light chain variable domain comprising an amino acid sequencewith at least 90%, at least 91%, at least 92%, at least 93%, at least94%, at least 95%, at least 96%, at least 97%, at least 98%, at least99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-19 or to the amino acid sequence of SEQ ID NO:79, wherein the light chain variable domain comprises the HVR-L1,HVR-L2, and HVR-L3 amino acid sequences of antibody S-60-19. In someembodiments, the anti-Sortilin antibody comprises a heavy chain variabledomain (VH) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a heavy chain variabledomain amino acid sequence of antibody S-60-19 or to the amino acidsequence of SEQ ID NO: 54 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the heavy chain variable domain amino acid sequence of antibodyS-60-19 or the amino acid sequence of SEQ ID NO: 54. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the heavy chain variable domain amino acidsequence of antibody S-60-19 or the amino acid sequence of SEQ ID NO:54. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VHsequence of antibody S-60-19 or of SEQ ID NO: 54, includingpost-translational modifications of that sequence. In a particularembodiment, the VH comprises one, two or three HVRs selected from: (a)the HVR-H1 amino acid sequence of antibody S-60-19, (b) the HVR-H2 aminoacid sequence of antibody S-60-19, and (c) the HVR-H3 amino acidsequence of antibody S-60-19. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a light chain variabledomain (VL) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a light chain variabledomain amino acid sequence of antibody S-60-19 or to the amino acidsequence of SEQ ID NO: 79 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the light chain variable domain amino acid sequence of antibodyS-60-19 or the amino acid sequence of SEQ ID NO: 79. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the light chain variable domain amino acidsequence of antibody S-60-19 or the amino acid sequence of SEQ ID NO:79. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VLsequence of antibody S-60-19 or of SEQ ID NO: 79, includingpost-translational modifications of that sequence. In a particularembodiment, the VL comprises one, two or three HVRs selected from: (a)the HVR-L1 amino acid sequence of antibody S-60-19, (b) the HVR-L2 aminoacid sequence of antibody S-60-19, and (c) the HVR-L3 amino acidsequence of antibody S-60-19.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain comprising the amino acid sequence of SEQ ID NO:105 or SEQ ID NO: 106. In some embodiments, anti-Sortilin antibodies ofthe present disclosure comprise a light chain comprising the amino acidsequence of SEQ ID NO: 133. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a heavy chain comprisingthe amino acid sequence of SEQ ID NO: 105 or SEQ ID NO: 106 and a lightchain comprising the amino acid sequence of SEQ ID NO: 133.

(11) S-60-24

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable domain and a light chain variabledomain, wherein the heavy chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a heavy chain variable domain amino acidsequence of antibody S-60-24 or to the amino acid sequence of SEQ ID NO:56; and/or the light chain variable domain comprises an amino acidsequence with at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-24 or to the amino acid sequence of SEQ ID NO:80. In some embodiments, anti-Sortilin antibodies of the presentdisclosure comprise a heavy chain variable domain comprising an aminoacid sequence with at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, at least 99%, or 100% identity to a heavy chain variable domainamino acid sequence of antibody S-60-24 or to the amino acid sequence ofSEQ ID NO: 56, wherein the heavy chain variable domain comprises theHVR-H1, HVR-H2, and HVR-H3 amino acid sequences of antibody S-60-24. Insome embodiments, anti-Sortilin antibodies of the present disclosurecomprise a light chain variable domain comprising an amino acid sequencewith at least 90%, at least 91%, at least 92%, at least 93%, at least94%, at least 95%, at least 96%, at least 97%, at least 98%, at least99%, or 100% identity to a light chain variable domain amino acidsequence of antibody S-60-24 or to the amino acid sequence of SEQ ID NO:80, wherein the light chain variable domain comprises the HVR-L1,HVR-L2, and HVR-L3 amino acid sequences of antibody S-60-24. In someembodiments, the anti-Sortilin antibody comprises a heavy chain variabledomain (VH) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a heavy chain variabledomain amino acid sequence of antibody S-60-24 or to the amino acidsequence of SEQ ID NO: 56 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the heavy chain variable domain amino acid sequence of antibodyS-60-24 or the amino acid sequence of SEQ ID NO: 56. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the heavy chain variable domain amino acidsequence of antibody S-60-24 or the amino acid sequence of SEQ ID NO:56. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VHsequence of antibody S-60-24 or of SEQ ID NO: 56, includingpost-translational modifications of that sequence. In a particularembodiment, the VH comprises one, two or three HVRs selected from: (a)the HVR-H1 amino acid sequence of antibody S-60-24, (b) the HVR-H2 aminoacid sequence of antibody S-60-24, and (c) the HVR-H3 amino acidsequence of antibody S-60-24. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a light chain variabledomain (VL) sequence having at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% identity to a light chain variabledomain amino acid sequence of antibody S-60-24 or to the amino acidsequence of SEQ ID NO: 80 and contains substitutions (e.g., conservativesubstitutions, insertions, or deletions relative to the referencesequence), but the anti-Sortilin antibody comprising that sequenceretains the ability to bind to Sortilin. In certain embodiments, a totalof 1 to 10 amino acids have been substituted, inserted, and/or deletedin the light chain variable domain amino acid sequence of antibodyS-60-24 or the amino acid sequence of SEQ ID NO: 80. In certainembodiments, a total of 1 to 5 amino acids have been substituted,inserted and/or deleted in the light chain variable domain amino acidsequence of antibody S-60-24 or the amino acid sequence of SEQ ID NO:80. In certain embodiments, substitutions, insertions, or deletionsoccur in regions outside the HVRs (i.e., in the FR regions). In someembodiments, the substitutions, insertions, or deletions occur in the FRregions. Optionally, the anti-Sortilin antibody comprises the VLsequence of antibody S-60-24 or of SEQ ID NO: 80, includingpost-translational modifications of that sequence. In a particularembodiment, the VL comprises one, two or three HVRs selected from: (a)the HVR-L1 amino acid sequence of antibody S-60-24, (b) the HVR-L2 aminoacid sequence of antibody S-60-24, and (c) the HVR-L3 amino acidsequence of antibody S-60-24.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain comprising the amino acid sequence of SEQ ID NO:137 or SEQ ID NO: 138. In some embodiments, anti-Sortilin antibodies ofthe present disclosure comprise a light chain comprising the amino acidsequence of SEQ ID NO: 134. In some embodiments, anti-Sortilinantibodies of the present disclosure comprise a heavy chain comprisingthe amino acid sequence of SEQ ID NO: 137 or SEQ ID NO: 138 and a lightchain comprising the amino acid sequence of SEQ ID NO: 134.

In some embodiments, an anti-Sortilin antibody of the present disclosurebinds essentially the same Sortilin epitope as an antibody comprisingthe heavy chain variable domain and the light chain variable domain ofan antibody selected from the group consisting of S-60-10, S-60-11,S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33I],S-60-16, S-60-18, S-60-19, and S-60-24.

In some embodiments, the anti-Sortilin antibody is anti-Sortilinmonoclonal antibody S-60-10. In some embodiments, the anti-Sortilinantibody is an isolated antibody which binds essentially the sameSortilin epitope as S-60-10. In some embodiments, the anti-Sortilinantibody is an isolated antibody comprising the heavy chain variableregion of monoclonal antibody S-60-10. In some embodiments, theanti-Sortilin antibody is an isolated antibody comprising the lightchain variable region of monoclonal antibody S-60-10. In someembodiments, the anti-Sortilin antibody is an isolated antibodycomprising the heavy chain variable region and the light chain variableregion of monoclonal antibody S-60-10.

In some embodiments, the anti-Sortilin antibody is anti-Sortilinmonoclonal antibody S-60-11. In some embodiments, the anti-Sortilinantibody is an isolated antibody which binds essentially the sameSortilin epitope as S-60-11. In some embodiments, the anti-Sortilinantibody is an isolated antibody comprising the heavy chain variableregion of monoclonal antibody S-60-11. In some embodiments, theanti-Sortilin antibody is an isolated antibody comprising the lightchain variable region of monoclonal antibody S-60-11. In someembodiments, the anti-Sortilin antibody is an isolated antibodycomprising the heavy chain variable region and the light chain variableregion of monoclonal antibody S-60-11.

In some embodiments, the anti-Sortilin antibody is anti-Sortilinmonoclonal antibody S-60-12. In some embodiments, the anti-Sortilinantibody is an isolated antibody which binds essentially the sameSortilin epitope as S-60-12. In some embodiments, the anti-Sortilinantibody is an isolated antibody comprising the heavy chain variableregion of monoclonal antibody S-60-12. In some embodiments, theanti-Sortilin antibody is an isolated antibody comprising the lightchain variable region of monoclonal antibody S-60-12. In someembodiments, the anti-Sortilin antibody is an isolated antibodycomprising the heavy chain variable region and the light chain variableregion of monoclonal antibody S-60-12.

In some embodiments, the anti-Sortilin antibody is anti-Sortilinmonoclonal antibody S-60-13. In some embodiments, the anti-Sortilinantibody is an isolated antibody which binds essentially the sameSortilin epitope as S-60-13. In some embodiments, the anti-Sortilinantibody is an isolated antibody comprising the heavy chain variableregion of monoclonal antibody S-60-13. In some embodiments, theanti-Sortilin antibody is an isolated antibody comprising the lightchain variable region of monoclonal antibody S-60-13. In someembodiments, the anti-Sortilin antibody is an isolated antibodycomprising the heavy chain variable region and the light chain variableregion of monoclonal antibody S-60-13.

In some embodiments, the anti-Sortilin antibody is anti-Sortilinmonoclonal antibody S-60-14. In some embodiments, the anti-Sortilinantibody is an isolated antibody which binds essentially the sameSortilin epitope as S-60-14. In some embodiments, the anti-Sortilinantibody is an isolated antibody comprising the heavy chain variableregion of monoclonal antibody S-60-14. In some embodiments, theanti-Sortilin antibody is an isolated antibody comprising the lightchain variable region of monoclonal antibody S-60-14. In someembodiments, the anti-Sortilin antibody is an isolated antibodycomprising the heavy chain variable region and the light chain variableregion of monoclonal antibody S-60-14.

In some embodiments, the anti-Sortilin antibody is anti-Sortilinmonoclonal antibody S-60-15. In some embodiments, the anti-Sortilinantibody is an isolated antibody which binds essentially the sameSortilin epitope as S-60-15. In some embodiments, the anti-Sortilinantibody is an isolated antibody comprising the heavy chain variableregion of monoclonal antibody S-60-15. In some embodiments, theanti-Sortilin antibody is an isolated antibody comprising the lightchain variable region of monoclonal antibody S-60-15. In someembodiments, the anti-Sortilin antibody is an isolated antibodycomprising the heavy chain variable region and the light chain variableregion of monoclonal antibody S-60-15.

In some embodiments, the anti-Sortilin antibody is anti-Sortilinmonoclonal antibody S-60-15.1. In some embodiments, the anti-Sortilinantibody is an isolated antibody which binds essentially the sameSortilin epitope as S-60-15.1. In some embodiments, the anti-Sortilinantibody is an isolated antibody comprising the heavy chain variableregion of monoclonal antibody S-60-15.1. In some embodiments, theanti-Sortilin antibody is an isolated antibody comprising the lightchain variable region of monoclonal antibody S-60-15.1. In someembodiments, the anti-Sortilin antibody is an isolated antibodycomprising the heavy chain variable region and the light chain variableregion of monoclonal antibody S-60-15.1.

In some embodiments, the anti-Sortilin antibody is anti-Sortilinmonoclonal antibody S-60-16. In some embodiments, the anti-Sortilinantibody is an isolated antibody which binds essentially the sameSortilin epitope as S-60-16. In some embodiments, the anti-Sortilinantibody is an isolated antibody comprising the heavy chain variableregion of monoclonal antibody S-60-16. In some embodiments, theanti-Sortilin antibody is an isolated antibody comprising the lightchain variable region of monoclonal antibody S-60-16. In someembodiments, the anti-Sortilin antibody is an isolated antibodycomprising the heavy chain variable region and the light chain variableregion of monoclonal antibody S-60-16.

In some embodiments, the anti-Sortilin antibody is anti-Sortilinmonoclonal antibody S-60-18. In some embodiments, the anti-Sortilinantibody is an isolated antibody which binds essentially the sameSortilin epitope as S-60-18. In some embodiments, the anti-Sortilinantibody is an isolated antibody comprising the heavy chain variableregion of monoclonal antibody S-60-18. In some embodiments, theanti-Sortilin antibody is an isolated antibody comprising the lightchain variable region of monoclonal antibody S-60-18. In someembodiments, the anti-Sortilin antibody is an isolated antibodycomprising the heavy chain variable region and the light chain variableregion of monoclonal antibody S-60-18.

In some embodiments, the anti-Sortilin antibody is anti-Sortilinmonoclonal antibody S-60-19. In some embodiments, the anti-Sortilinantibody is an isolated antibody which binds essentially the sameSortilin epitope as S-60-19. In some embodiments, the anti-Sortilinantibody is an isolated antibody comprising the heavy chain variableregion of monoclonal antibody S-60-19. In some embodiments, theanti-Sortilin antibody is an isolated antibody comprising the lightchain variable region of monoclonal antibody S-60-19. In someembodiments, the anti-Sortilin antibody is an isolated antibodycomprising the heavy chain variable region and the light chain variableregion of monoclonal antibody S-60-19.

In some embodiments, the anti-Sortilin antibody is anti-Sortilinmonoclonal antibody S-60-24. In some embodiments, the anti-Sortilinantibody is an isolated antibody which binds essentially the sameSortilin epitope as S-60-24. In some embodiments, the anti-Sortilinantibody is an isolated antibody comprising the heavy chain variableregion of monoclonal antibody S-60-24. In some embodiments, theanti-Sortilin antibody is an isolated antibody comprising the lightchain variable region of monoclonal antibody S-60-24. In someembodiments, the anti-Sortilin antibody is an isolated antibodycomprising the heavy chain variable region and the light chain variableregion of monoclonal antibody S-60-24.

In certain embodiments, the anti-Sortilin antibody is an antagonistantibody. In certain embodiments, the anti-Sortilin antibody is anagonist antibody. In some embodiments, anti-Sortilin antibodies of thepresent disclosure are of the IgG class the IgM class, or the IgA class.In some embodiments, anti-Sortilin antibodies of the present disclosureare of the IgG class and have an IgG1, IgG2, IgG3, or IgG4 isotype.

Additional anti-Sortilinantibodies, e.g., antibodies that specificallybind to a Sortilin protein of the present disclosure, may be identified,screened, and/or characterized for their physical/chemical propertiesand/or biological activities by various assays known in the art.

Certain aspects of the present disclosure relate to the use of two ormore anti-Sortilin antibodies that when utilized together displayadditive or synergistic effects, as compared to utilization of acorresponding single anti-Sortilin antibody.

In some embodiments, an anti-Sortilin antibody of the present disclosureis an antibody fragment that binds to a human Sortilin protein.

In some embodiments, an anti-Sortilin antibody of the present disclosureis an antibody fragment that binds to one or more human proteinsselected from the group consisting of human Sortilin, a naturallyoccurring variant of human Sortilin, and a disease variant of humanSortilin.

In some embodiments, an anti-Sortilin antibody of the present disclosureis antibody fragment, wherein the antibody fragment is an Fab, Fab′,Fab′-SH, F(ab′)2, Fv, or scFv fragment.

Antibody Frameworks

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable region comprising one or more (e.g., oneor more, two or more, three or more, or all four) framework regionsselected from VH FR1, VH FR2, VH FR3, and VH FR4 (as shown in Tables20-23). In some embodiments, the VH FR1 comprises a sequence ofQVQLQESGPGLVKPSETLSL TCAVSG (SEQ ID NO: 35). In some embodiments, the VHFR2 comprises a sequence of WIRQPPGKGLEWIG (SEQ ID NO: 36). In someembodiments, the VH FR3 comprises the sequence according to Formula VI:X₁VTISVDTSKNQFSLX₂LSSVTAADTAVYYC (SEQ ID NO: 39), wherein X₁ is Q or R,and X₂ is E or K. In some embodiments, VH FR3 comprises a sequenceselected from the group consisting of SEQ ID NOs: 37-38. In someembodiments, VH FR4 comprises a sequence of WGQGTTVTVSS (SEQ ID NO: 40).In some embodiments, an antibody comprises a heavy chain variable regioncomprising a VH FR1 comprising the sequence of SEQ ID NO: 35, a VH FR2comprising the sequence of SEQ ID NO: 36, a VH FR3 according to FormulaVI, and a VH FR4 comprising the sequence of SEQ ID NO: 40.

In some embodiments, an antibody comprises a heavy chain variable regioncomprising a VH FR1 comprising the sequence of SEQ ID NO: 35, a VH FR2comprising the sequence of SEQ ID NO: 36, a VH FR3 comprising thesequence selected from SEQ ID NOs: 37-38, and a VH FR4 comprising thesequence of SEQ ID NO: 40.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable region comprising a VH FR1, a VH FR2, aVH FR3, and VH FR4 of antibody S-60-10, S-60-11, S-60-12, S-60-13,S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S],S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H],S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E],S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14[N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L],S-60-16; S-60-18, S-60-19, or S-60-24 (as shown in Tables 20-23).

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a light chain variable region comprising one or more (e.g., oneor more, two or more, three or more, or all four) framework regionsselected from VL FR1, VL FR2, VL FR3, and VL FR4 (as shown in Tables24-27).

In some embodiments, the VL FR1 comprises a sequence according toFormula VII: DIVMTQSPLSLPVTPGX₁X₂ASISC (SEQ ID NO: 44), wherein X₁ is Eor G, and X₂ is P or S. In some embodiments, VL FR1 comprises a sequenceselected from the group consisting of SEQ ID NOs: 41-43. In someembodiments, the VL FR2 comprises a sequence according to Formula VIII:WYLQKPGQX₁PQLLIY (SEQ ID NO: 47), wherein X₁ is S or P. In someembodiments, VL FR2 comprises a sequence selected from the groupconsisting of SEQ ID NOs: 45-46. In some embodiments, the VL FR3comprises a sequence according to Formula IX: GVPDRX₁SGSGSGTDFTLKISRX₂EAEDVGX₃YYC (SEQ ID NO: 52), wherein X₁ is F or L, X₂ is A orV, and X₃ is V or A. In some embodiments, VL FR3 comprises a sequenceselected from the group consisting of SEQ ID NOs: 48-51. In someembodiments, the VL FR4 comprises a sequence of FGGGTKVEIK (SEQ ID NO:53). In some embodiments, anti-Sortilin antibodies of the presentdisclosure comprise a light chain variable region comprising a VL FR1comprising the sequence according to Formula VII, a VL FR2 comprisingthe sequence according to Formula VIII, a VL FR3 comprising the sequenceaccording to Formula IX, and a VL FR4 comprising the sequence of SEQ IDNO: 53.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a light chain variable region comprising a VL FR1 comprisingthe sequence selected from SEQ ID NOs: 41-43, a VL FR2 comprising thesequence selected from SEQ ID NOs: 45-46, a VL FR3 comprising thesequence selected from SEQ ID NOs: 48-51, and a VL FR4 comprising thesequence of SEQ ID NO: 53.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a light chain variable region comprising a VL FR1, a VL FR2, aVL FR3, and VL FR4 of antibody S-60-10, S-60-11, S-60-12, S-60-13,S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S],S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H],S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E],S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14[N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L],S-60-16; S-60-18, S-60-19, or S-60-24 (as shown in Tables 24-27).

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable region comprising one or more (e.g., oneor more, two or more, three or more, or all four) framework regionsselected from VH FR1, VH FR2, VH FR3, and VH FR4 (as shown in Tables20-23), and a light chain variable region comprising one or more (e.g.,one or more, two or more, three or more, or all four) framework regionsselected from VL FR1, VL FR2, VL FR3, and VL FR4 (as shown in Tables24-27). In some embodiments, anti-Sortilin antibodies of the presentdisclosure comprise a heavy chain variable region comprising a VH FR1comprising the sequence of SEQ ID NO: 35, a VH FR2 comprising thesequence of SEQ ID NO: 36, a VH FR3 according to Formula VI, and a VHFR4 comprising the sequence of SEQ ID NO: 40; and a light chain variableregion comprising a VL FR1 comprising the sequence according to FormulaVII, a VL FR2 comprising the sequence according to Formula VIII, a VLFR3 comprising the sequence according to Formula IX, and a VL FR4comprising the sequence of SEQ ID NO: 53. In some embodiments,anti-Sortilin antibodies of the present disclosure comprise a heavychain variable region comprising a VH FR1 comprising the sequence of SEQID NO: 35, a VH FR2 comprising the sequence of SEQ ID NO: 36, a VH FR3comprising the sequence selected from SEQ ID NOs: 37-38, and a VH FR4comprising the sequence of SEQ ID NO: 40; a light chain variable regioncomprising a VL FR1 comprising the sequence selected from SEQ ID NOs:41-43, a VL FR2 comprising the sequence selected from SEQ ID NOs: 45-46,a VL FR3 comprising the sequence selected from SEQ ID NOs: 48-51, and aVL FR4 comprising the sequence of SEQ ID NO: 53.

In some embodiments, anti-Sortilin antibodies of the present disclosurecomprise a heavy chain variable region comprising a VH FR1, a VH FR2, aVH FR3, and VH FR4 of antibody S-60-10, S-60-11, S-60-12, S-60-13,S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S],S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H],S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E],S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14[N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L],S-60-16; S-60-18, S-60-19, or S-60-24 (as shown in Tables 20-23), and alight chain variable region comprising a VL FR1, a VL FR2, a VL FR3, andVL FR4 of antibody S-60-10, S-60-11, S-60-12, S-60-13, S-60-14, S-60-15[N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G],S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K],S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11[N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V],S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16;S-60-18, S-60-19, or S-60-24 (as shown in Tables 24-27).

Anti-Sortilin Antibody Activities

In certain aspects of any of the anti-Sortilin antibodies, anti-Sortilinantibodies of the present disclosure can inhibit one or more activitiesof a Sortilin protein, including, but not limited to, decreasingcellular levels of Sortilin (e.g., cell surface levels of Sortilin,intracellular levels of Sortilin, and/or total levels of Sortilin);increasing Progranulin levels (e.g., extracellular levels of Progranulinand/or cellular levels of Progranulin); and inhibiting the interaction(e.g., binding) between Progranulin and Sortilin. As contemplatedherein, anti-Sortilin antibodies of the present disclosure may inhibitadditional activities of a Sortilin protein, including but not limitedto inhibiting interaction (e.g., binding) with one or more ofpro-neurotrophins of the present disclosure (pro-neurotrophin-3,pro-neurotrophin-4/5, pro-NGF, pro-BDNF, etc.), neurotrophins of thepresent disclosure (neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.),neurotensin, p75, Sortilin propeptide (Sort-pro), amyloid precursorprotein (APP), the A beta peptide, lipoprotein lipase (LpL),apolipoprotein AV (APOA5), apolipoprotein E (APOE), and receptorassociated protein (RAP), decreasing secretion of PCSK9, decreasingproduction of beta amyloid peptide.

In certain embodiments, the present disclosure provides an anti-Sortilinantibody, wherein (a) the anti-Sortilin antibody increases extracellularlevels of Progranulin, decreases cellular levels of Sortilin, inhibitsinteraction between Sortilin and Progranulin, or any combinationthereof; (b) the anti-Sortilin antibody decreases cell surface levels ofSortilin, increases extracellular levels of Progranulin, inhibitsinteraction between Sortilin and Progranulin, or any combinationthereof; (c) the anti-Sortilin antibody decreases cell surface levels ofSortilin, decreases intracellular levels of Sortilin, decreases totallevels of Sortilin, or any combination thereof; (d) the anti-Sortilinantibody induces Sortilin degradation, Sortilin cleavage, Sortilininternalization, Sortilin down regulation, or any combination thereof;(e) the anti-Sortilin antibody decreases cellular levels of Sortilin andinhibits the interaction between Sortilin and Progranulin; (f) theanti-Sortilin antibody decreases cellular levels of Sortilin andincreases cellular levels of Progranulin; and/or (g) the anti-Sortilinantibody increases the effective concentration of Progranulin.

In certain embodiments, the present disclosure provides an anti-Sortilinantibody, wherein the anti-Sortilin antibody decreases cell surfacelevels of Sortilin, increases extracellular levels of Progranulin,inhibits interaction between Sortilin and Progranulin, or anycombination thereof.

In some embodiments, an anti-Sortilin antibody of the present disclosure(a) reduces cell surface levels of Sortilin with a half maximaleffective concentration (EC₅₀) that is less than 150 pM, as measured byflow cytometry; (b) reduces cell surface levels of Sortilin by more thanabout 50% at 1.25 nM IgG, by more than about 80% at 0.63 nM IgG, or bymore than about 69% at 150 nM IgG relative to control, as measured byflow cytometry; increases Progranulin secretion by more than about 1.13fold over control at 0.63 nM IgG, or by more than about 1.22 fold overcontrol at 50 nM IgG, as measured by standard ELISA; blocks binding ofProgranulin to Sortilin with a half maximal effective concentration(EC₅₀) that is less than 0.325 nM, as measured by flow cytometry; (e)blocks binding of Progranulin to Sortilin by more than about 88% at 50nM IgG, or by more than about 27.5% at 150 nM IgG relative to control,as measured by flow cytometry; or (f) any combination thereof (see,e.g., Examples 2-4).

In some embodiments, an anti-Sortilin antibody of the present disclosure(a) reduces cell surface levels of Sortilin with a half maximaleffective concentration (EC₅₀) that is less than 681 pM, as measured byflow cytometry; (b) reduces cell surface levels of Sortilin by more thanabout 40% at 1.25 nM IgG, by more than about 29% at 0.6 nM IgG, or bymore than about 62% at 150 nM IgG relative to control, as measured byflow cytometry; (c) increases Progranulin secretion by more than about1.11 fold over control at 0.63 nM IgG, or by more than about 1.75 foldover control at 50 nM IgG, as measured by standard ELISA; (d) blocksbinding of Progranulin to Sortilin with a half maximal effectiveconcentration (EC₅₀) that is less than 0.751 nM, as measured by flowcytometry; (e) blocks binding of Progranulin to Sortilin by more thanabout 90% at 50 nM IgG, or by more than about 95% at 150 nM IgG relativeto control, as measured by flow cytometry; or (f) any combinationthereof (see, e.g., Examples 2-4).

Decreasing Sortilin Levels

In some embodiments, anti-Sortilin antibodies of the present disclosurebind to a Sortilin protein of the present disclosure expressed on thesurface of a cell and modulate (e.g., induce or inhibit) one or moreSortilin activities of the present disclosure after binding to thesurface-expressed Sortilin protein.

In some embodiments, anti-Sortilin antibodies of the present disclosuredecrease cellular levels of Sortilin in vitro. In some embodiments,anti-Sortilin antibodies of the present disclosure may decrease cellularlevels of Sortilin in vivo (e.g., in the brain, and/or peripheral organsof an individual). In some embodiments, a decrease in cellular levels ofSortilin comprises a decrease in cell surface levels of Sortilin. Asused herein, an anti-Sortilin antibody decreases cell surface levels ofSortilin if it induces a decrease at saturating antibody concentrations(e.g., 0.6 nM, 0.63 nM, 1.25 nM, 50 nM or 150 nM) and/or relative to acontrol antibody (e.g. an anti-Sortilin antibody having a heavy chainvariable region and a light chain variable region corresponding to S-60)in cell surface levels of Sortilin as measured by any in vitrocell-based assays or suitable in vivo model described herein or known inthe art. In some embodiments, a decrease in cellular levels of Sortilincomprises a decrease in intracellular levels of Sortilin. Ascontemplated herein, an anti-Sortilin antibody decreases intracellularlevels of Sortilin if it induces a decrease at saturating antibodyconcentrations and/or relative to a control antibody (e.g. ananti-Sortilin antibody having a heavy chain variable region and a lightchain variable region corresponding to S-60) in intracellular levels ofSortilin as measured by any in vitro cell-based assays or suitable invivo model described herein or known in the art. In some embodiments, adecrease in cellular levels of Sortilin comprises a decrease in totallevels of Sortilin. As contemplated herein, an anti-Sortilin antibodydecreases total levels of Sortilin if it induces a decrease atsaturating antibody concentrations and/or relative to a control antibody(e.g. an anti-Sortilin antibody having a heavy chain variable region anda light chain variable region corresponding to S-60) in total levels ofSortilin as measured by any in vitro cell-based assays or suitable invivo model described herein or known in the art.

As used herein, levels of Sortilin may refer to expression levels of thegene encoding Sortilin; to expression levels of one or more transcriptsencoding Sortilin; to expression levels of Sortilin protein; and/or tothe amount of Sortilin protein present within cells and/or on the cellsurface. Any methods known in the art for measuring levels of geneexpression, transcription, translation, and/or protein abundance orlocalization may be used to determine the levels of Sortilin.

Cellular levels of Sortilin may refer to, without limitation, cellsurface levels of Sortilin, intracellular levels of Sortilin, and totallevels of Sortilin. In some embodiments, a decrease in cellular levelsof Sortilin comprises decrease in cell surface levels of Sortilin. Insome embodiments, anti-Sortilin antibodies of the present disclosurethat decrease cellular levels of Sortilin (e.g., cell surface levels ofSortilin) have one or more of the following characteristics: (1)inhibits or reduces one or more Sortilin activities; (2) the ability toinhibit or reduce binding of a Sortilin to one or more of its ligands;(3) the ability to reduce Sortilin expression in Sortilin-expressingcells; (4) the ability to interact, bind, or recognize a Sortilinprotein; (5) the ability to specifically interact with or bind to aSortilin protein; and (6) the ability to treat, ameliorate, or preventany aspect of a disease or disorder described or contemplated herein.

In some embodiments, an isolated anti-Sortilin antibody of the presentdisclosure induces downregulation of Sortilin. In some embodiments, anisolated anti-Sortilin antibody of the present disclosure inducescleavage of Sortilin. In some embodiments, an isolated anti-Sortilinantibody of the present disclosure induces internalization of Sortilin.In some embodiments, an isolated anti-Sortilin antibody of the presentdisclosure induces shedding of Sortilin. In some embodiments, anisolated anti-Sortilin antibody of the present disclosure inducesdegradation of Sortilin. In some embodiments, an isolated anti-Sortilinantibody of the present disclosure induces desensitization of Sortilin.In some embodiments, an isolated anti-Sortilin antibody of the presentdisclosure acts as a ligand mimetic to transiently activate Sortilin. Insome embodiments, an isolated anti-Sortilin antibody of the presentdisclosure acts as a ligand mimetic and transiently activates Sortilinbefore inducing a decrease in cellular levels of Sortilin and/orinhibition of interaction (e.g., binding) between Sortilin and one ormore Sortilin ligands. In some embodiments, an isolated anti-Sortilinantibody of the present disclosure acts as a ligand mimetic andtransiently activates Sortilin before inducing degradation of Sortilin.In some embodiments, an isolated anti-Sortilin antibody of the presentdisclosure acts as a ligand mimetic and transiently activates Sortilinbefore inducing cleavage of Sortilin. In some embodiments, an isolatedanti-Sortilin antibody of the present disclosure acts as a ligandmimetic and transiently activates Sortilin before inducinginternalization of Sortilin. In some embodiments, an isolatedanti-Sortilin antibody of the present disclosure acts as a ligandmimetic and transiently activates Sortilin before inducing shedding ofSortilin. In some embodiments, an isolated anti-Sortilin antibody of thepresent disclosure acts as a ligand mimetic and transiently activatesSortilin before inducing downregulation of Sortilin expression. In someembodiments, an isolated anti-Sortilin antibody of the presentdisclosure acts as a ligand mimetic and transiently activates Sortilinbefore inducing desensitization of Sortilin.

In certain embodiments, anti-Sortilin antibodies of the presentdisclosure may decrease cellular levels of Sortilin (e.g., cell surfacelevels of Sortilin, intracellular levels of Sortilin, and/or totallevels of Sortilin) by inducing Sortilin degradation. Accordingly, insome embodiments, anti-Sortilin antibodies of the present disclosureinduce Sortilin degradation.

Anti-Sortilin antibodies of the present disclosure may decrease cellularlevels (e.g., cell surface levels) of Sortilin with a half-maximaleffective concentration (EC₅₀) (e.g., when measured in vitro) in thepicomolar range. In certain embodiments, the EC₅₀ of the antibody isless than about 680.9 pM. In certain embodiments, the EC₅₀ of theantibody is about 72.58 pM to about 680.9 nM. In certain embodiments,the EC₅₀ of the antibody is about 103.6 pM to about 680.9 nM. In certainembodiments, the EC₅₀ of the antibody is less than about 600 pM, 500 pM,400 pM, 300 pM, 200 pM, 100 pM, 50 pM, 40 pM, 30 pM, 20 pM, 10 pM, 1 pM,or 0.5 pM.

In some embodiments, the EC₅₀ of the antibody is less than about orequal to about 675 pM, 650 pM, 625 pM, 600 pM, 575 pM, 550 pM, 525 pM,500 pM, 475 pM, 450 pM, 425 pM, 400 pM, 375 pM, 350 pM, 325 pM, 300 pM,275 pM, 250 pM, 225 pM, 200 pM, 175 pM, 150 pM, 125 pM, 100 pM, 90 pM,80 pM, 70 pM, 60 pM, 50 pM, 40 pM, 30 pM, 20 pM, 10 pM, 9 pM, 8 pM, 7pM, 6 pM, 5 pM, 4 pM, 3 pM, 2 pM, 1 pM, or 0.5 pM.

In some embodiments, the EC₅₀ of the antibody is less than about 680.9pM. In some embodiments, the EC₅₀ of the antibody is greater than aboutor equal to about 0.1 pM, 0.5 pM, 1 pM, 10 pM, 20 pM, 30 pM, 40 pM, 50pM, 60 pM, 70 pM, 80 pM, 90 pM, 100 pM, 125 pM, 150 pM, 175 pM, 200 pM,225 pM, 250 pM, 275 pM, 300 pM, 325 pM, 350 pM, 375 pM, 400 pM, 425 pM,450 pM, 475 pM, 500 pM, 525 pM, 550 pM, 575 pM, 600 pM, 625 pM, 650 pM,675 pM. That is, the EC₅₀ of the antibody can be any of a range havingan upper limit of about 675 pM, 650 nM, 650 pM, 625 pM, 600 pM, 575 pM,550 pM, 525 pM, 500 pM, 475 pM, 450 pM, 425 pM, 400 pM, 375 pM, 350 pM,325 pM, 300 pM, 275 pM, 250 pM, 225 pM, 200 pM, 175 pM, 150 pM, 125 pM,100 pM, 90 pM, 80 pM, 70 pM, 60 pM, 50 pM, 40 pM, 30 pM, 20 pM, 10 pM, 1pM, or 0.5 pM, and an independently selected lower limit of about 0.1pM, 0.5 pM, 1 pM, 10 pM, 20 pM, 30 pM, 40 pM, 50 pM, 60 pM, 70 pM, 80pM, 90 pM, 100 pM, 125 pM, 150 pM, 175 pM, 200 pM, 225 pM, 250 pM, 275pM, 300 pM, 325 pM, 350 pM, 375 pM, 400 pM, 425 pM, 450 pM, 475 pM, 500pM, 525 pM, 550 pM, 575 pM, 600 pM, 625 pM, 650 pM, or 675 pM, whereinthe lower limit is less than the upper limit. In some embodiments, theEC₅₀ of the antibody is any of about 1 pM, 2 pM, 3 pM, 4 pM, 5 pM, 6 pM,7 pM, 8 pM, 9 pM, 10 pM, 15 pM, 20 pM, 25 pM, 30 pM, 35 pM, 40 pM, 45pM, 50 pM, 55 pM, 60 pM, 65 pM, 70 pM, 75 pM, 80 pM, 85 pM, 90 pM, 95pM, 100 pM, 105 pM, 110 pM, 115 pM, 120 pM, 125 pM, 130 pM, 135 pM, 140pM, 145 pM, 150 pM, 155 pM, 160 pM, 165 pM, 170 pM, 175 pM, 180 pM, 185pM, 190 pM, 195 pM, or 200 pM.

In some embodiments, an anti-Sortilin antibody of the present disclosurereduces cell surface levels of Sortilin with a half maximal effectiveconcentration (EC₅₀) that is less than 150 pM, as measured by flowcytometry. In some embodiments, the EC₅₀ of an anti-Sortilin antibody ofthe present disclosure is about 103.6 pM. In some embodiments, the EC₅₀of an anti-Sortilin antibody of the present disclosure is about 72.58pM.

In some embodiments, an anti-Sortilin antibody of the present disclosurereduces cell surface levels of Sortilin by more than about 40% at 1.25nM IgG or by more than about 80% at 0.63 nM IgG, as measured by flowcytometry. In some embodiments, an anti-Sortilin antibody of the presentdisclosure reduces cell surface levels of Sortilin by about 60.92% at1.25 nM IgG, as measured by flow cytometry. In some embodiments, ananti-Sortilin antibody of the present disclosure reduces cell surfacelevels of Sortilin by about 69.3% at 150 nM IgG, as measured by flowcytometry. In some embodiments, an anti-Sortilin antibody of the presentdisclosure reduces cell surface levels of Sortilin by about 70.3% at 150nM IgG, as measured by flow cytometry.

Various methods of measuring antibody EC₅₀ values are known in the art,including, for example, by flow cytometry (See e.g., Example 2). In someembodiments, the EC₅₀ is measured in vitro using cells engineered toexpress human Sortilin. In some embodiments, the EC₅₀ is measured at atemperature of approximately 4° C. In some embodiments, the EC₅₀ ismeasured at a temperature of approximately 25° C. In some embodiments,the EC₅₀ is measured at a temperature of approximately 35° C. In someembodiments, the EC₅₀ is measured at a temperature of approximately 37°C. In some embodiments, the EC₅₀ is determined using a monovalentantibody (e.g., a Fab) or a full-length antibody in a monovalent form.In some embodiments, the EC₅₀ is determined using antibodies containingconstant regions that demonstrate enhanced Fc receptor binding. In someembodiments, the EC₅₀ is determined using antibodies containing constantregions that demonstrate reduced Fc receptor binding.

In some embodiments, anti-Sortilin antibodies of the present disclosurehave higher potencies in reducing cell surface levels of Sortilinrelative to a control antibody (e.g. an anti-Sortilin antibody having aheavy chain variable region and a light chain variable regioncorresponding to S-60). In some embodiments, anti-Sortilin antibodies ofthe present disclosure decrease cellular levels (e.g., cell surfacelevels) of Sortilin with a lower EC₅₀ (e.g., as measured in vitro) thana control antibody (e.g. an anti-Sortilin antibody having a heavy chainvariable region and a light chain variable region corresponding toS-60). In some embodiments, anti-Sortilin antibodies of the presentdisclosure decrease cellular levels (e.g., cell surface levels) ofSortilin with an EC₅₀ that is at least about 5%, at least about 10%, atleast about 15%, at least about 20%, at least about 25%, at least about30%, at least about 35%, at least about 40%, at least about 45%, atleast about 50%, at least about 55%, at least about 60%, at least about65%, at least about 70%, at least about 75%, at least about 80%, atleast about 85%, at least about 90%, at least about 95%, or at leastabout 99% lower than the EC₅₀ of a control antibody (e.g. ananti-Sortilin antibody having a heavy chain variable region and a lightchain variable region corresponding to S-60). In some embodiments,anti-Sortilin antibodies of the present disclosure decrease cellularlevels (e.g., cell surface levels) of Sortilin with an EC₅₀ that is atleast about 1-fold, at least about 1.1-fold, at least about 1.5-fold, atleast about 2-fold, at least about 3-fold, at least about 4-fold, atleast about 5-fold, at least about 6-fold, at least about 7-fold, atleast about 8-fold, at least about 9-fold, at least about 10-fold, atleast about 12.5-fold, at least about 15-fold, at least about 17.5-fold,at least about 20-fold, at least about 22.5-fold, at least about25-fold, at least about 27.5-fold, at least about 30-fold, at leastabout 50-fold, or at least about 100-fold lower than the EC₅₀ of acontrol antibody (e.g. an anti-Sortilin antibody having a heavy chainvariable region and a light chain variable region corresponding toS-60).

In some embodiments, anti-Sortilin antibodies of the present disclosurehave an EC₅₀ that is at least 1.5-fold lower than control antibody (e.g.an anti-Sortilin antibody having a heavy chain variable region and alight chain variable region corresponding to S-60). In some embodiments,anti-Sortilin antibodies of the present disclosure have an EC₅₀ that isat least 1.1-fold lower than control antibody (e.g. an anti-Sortilinantibody having a heavy chain variable region and a light chain variableregion corresponding to S-60).

In some embodiments, an anti-Sortilin antibody of the present disclosure(a) reduces cell surface levels of Sortilin with a half maximaleffective concentration (EC₅₀) that is less than 681 pM, as measured byflow cytometry; (b) reduces cell surface levels of Sortilin by more thanabout 40% at 1.25 nM IgG, by more than about 29% at 0.6 nM IgG, or bymore than about 62% at 150 nM IgG relative to control, as measured byflow cytometry; (c) increases Progranulin secretion by more than about1.11 fold over control at 0.63 nM IgG, or by more than about 1.75 foldover control at 50 nM IgG, as measured by standard ELISA; (d) blocksbinding of Progranulin to Sortilin with a half maximal effectiveconcentration (EC₅₀) that is less than 0.751 nM, as measured by flowcytometry; (e) blocks binding of Progranulin to Sortilin by more thanabout 90% at 50 nM IgG, or by more than about 95% at 150 nM IgG relativeto control, as measured by flow cytometry; or (f) any combinationthereof (see, e.g., Examples 2-4).

Increasing Progranulin Levels

In some embodiments, anti-Sortilin antibodies of the present disclosureincrease extracellular levels of Progranulin in vitro. In someembodiments, anti-Sortilin antibodies of the present disclosure mayincrease cellular levels of Progranulin or in vivo (e.g., in the brain,blood, and/or peripheral organs of an individual). As used herein, ananti-Sortilin antibody increases extracellular levels of Progranulin ifit induces an increase at saturating antibody concentrations (e.g., 0.6nM, 0.63 nM, 1.25 nM, 50 nM or 150 nM) and/or relative to a controlantibody (e.g. an anti-Sortilin antibody having a heavy chain variableregion and a light chain variable region corresponding to S-60) inextracellular levels of Progranulin as measured by any in vitrocell-based assays or in tissue-based (such as brain tissue-based) assaysdescribed herein or known in the art. As contemplated herein, ananti-Sortilin antibody increases cellular levels of Progranulin if itinduces an increase at saturating antibody concentrations (e.g., 0.6 nM,0.63 nM, 1.25 nM, 50 nM or 150 nM) and/or relative to a control antibody(e.g. an anti-Sortilin antibody having a heavy chain variable region anda light chain variable region corresponding to S-60) in cellular levelsof Progranulin as measured by any in vitro cell-based assays or intissue-based (such as brain tissue-based) assays described herein orknown in the art.

As used herein, levels of Progranulin may refer to expression levels ofthe gene encoding Progranulin; to expression levels of one or moretranscripts encoding Progranulin; to expression levels of Progranulinprotein; and/or to the amount of Progranulin protein secreted from cellsand/or present within cells. Any methods known in the art for measuringlevels of gene expression, transcription, translation, proteinabundance, protein secretion, and/or protein localization may used todetermine the levels of Progranulin.

As used herein, Progranulin levels may refer to, without limitation,extracellular levels of Progranulin, intracellular levels ofProgranulin, and total levels of Progranulin. In some embodiments, anincrease in levels of Progranulin comprises an increase in extracellularlevels of Progranulin.

In some embodiments, an anti-Sortilin antibody of the present disclosureincreases Progranulin secretion by more than about 1.11 fold overcontrol at 0.63 nM IgG, as measured by standard ELISA. In someembodiments, an anti-Sortilin antibody of the present disclosureincreases Progranulin secretion by about 1.42 fold over control at 0.63nM IgG, as measured by standard ELISA. In some embodiments, ananti-Sortilin antibody of the present disclosure increases Progranulinsecretion by more than about 1.75 fold over control at 50 nM IgG, asmeasured by standard ELISA. In some embodiments, an anti-Sortilinantibody of the present disclosure increases Progranulin secretion byabout 1.97 fold over control at 50 nM IgG, as measured by standardELISA. In some embodiments, an anti-Sortilin antibody of the presentdisclosure increases Progranulin secretion by about 2.29 fold overcontrol at 50 nM IgG, as measured by standard ELISA.

Various methods of measuring Progranulin secretion are known in the art,including, for example, by ELISA (See e.g., Examples 2 and 4). In someembodiments, the EC₅₀ is measured in vitro using cells expressing humanSortilin. In some embodiments, Progranulin secretion is determined usinga monovalent antibody (e.g., a Fab) or a full-length antibody in amonovalent form. In some embodiments, Progranulin secretion isdetermined using antibodies containing constant regions that demonstrateenhanced Fc receptor binding. In some embodiments, Progranulin secretionis determined using antibodies containing constant regions thatdemonstrate reduced Fc receptor binding.

In some embodiments, anti-Sortilin antibodies of the present disclosurehave higher potencies in increasing levels of Progranulin relative to acontrol antibody (e.g. an anti-Sortilin antibody having a heavy chainvariable region and a light chain variable region corresponding toS-60). In some embodiments, anti-Sortilin antibodies of the presentdisclosure increase levels (e.g., extracellular levels) of Progranulinwith a lower EC₅₀ (e.g., as measured in vitro) than a control antibody(e.g. an anti-Sortilin antibody having a heavy chain variable region anda light chain variable region corresponding to S-60). In someembodiments, anti-Sortilin antibodies of the present disclosure increaselevels (e.g., extracellular levels) of Progranulin by at least about 5%,at least about 10%, at least about 15%, at least about 20%, at leastabout 25%, at least about 30%, at least about 35%, at least about 40%,at least about 45%, at least about 50%, at least about 55%, at leastabout 60%, at least about 65%, at least about 70%, at least about 75%,at least about 80%, at least about 85%, at least about 90%, at leastabout 95%, or at least about 99% than a control antibody (e.g. ananti-Sortilin antibody having a heavy chain variable region and a lightchain variable region corresponding to S-60). In some embodiments,anti-Sortilin antibodies of the present disclosure increase levels(e.g., extracellular levels) of Progranulin by about 1-fold, at leastabout 1.1-fold, at least about 1.5-fold, at least about 2-fold, at leastabout 3-fold, at least about 4-fold, at least about 5-fold, at leastabout 6-fold, at least about 7-fold, at least about 8-fold, at leastabout 9-fold, at least about 10-fold, at least about 12.5-fold, at leastabout 15-fold, at least about 17.5-fold, at least about 20-fold, atleast about 22.5-fold, at least about 25-fold, at least about 27.5-fold,at least about 30-fold, at least about 50-fold, or at least about100-fold higher than a control antibody (e.g. an anti-Sortilin antibodyhaving a heavy chain variable region and a light chain variable regioncorresponding to S-60).

In some embodiments, anti-Sortilin antibodies of the present disclosureincrease Progranulin levels by about 1.1-fold higher than a controlantibody (e.g. an anti-Sortilin antibody having a heavy chain variableregion and a light chain variable region corresponding to S-60). In someembodiments, anti-Sortilin antibodies of the present disclosure increaseProgranulin levels by about 1.3-fold higher than a control antibody(e.g. an anti-Sortilin antibody having a heavy chain variable region anda light chain variable region corresponding to S-60).

In some embodiments, anti-Sortilin antibodies of the present disclosureincrease the effective concentration of Progranulin. The effectiveconcentration of Progranulin refers to the concentration of Progranulinin plasma or cerebrospinal fluid. In some embodiments, an increase inthe effective concentration of Progranulin is an increase of greaterthan 1.5 fold. In some embodiments, the effective concentration ofProgranulin is increased for 7-28 days.

Decreasing Interaction Between Sortilin and Binding Partner

In certain embodiments, the anti-Sortilin antibodies inhibit interaction(e.g., binding) between a Sortilin protein of the present disclosure andone or more proteins selected from Progranulin, a pro-neurotrophin, aneurotrophin, pro-neurotrophin-3, neurotrophin-3, pro-neurotrophin-4/5,neurotrophin-4/5, pro-nerve growth factor (pro-NGF), nerve growth factor(NGF), pro-brain-derived neurotrophic factor (pro-BDNF), brain-derivedneurotrophic factor (BDNF), neurotensin, p75, Sortilin propeptide(Sort-pro), amyloid precursor protein (APP), the A beta peptide,lipoprotein lipase (LpL), apolipoprotein AV (APOA5), apolipoprotein E(APOE), PCSK9, and receptor associated protein (RAP), and/or naturallyoccurring variants. In a specific embodiment, anti-Sortilin antibodiesof the present disclosure inhibit interaction (e.g., binding) between aSortilin protein of the present disclosure and Progranulin.

In some embodiments, the anti-Sortilin antibodies may inhibitinteraction (e.g., binding) between a Sortilin protein of the presentdisclosure and a neurotrophin of the present disclosure, such as apro-neurotrophin, pro-neurotrophin-3, pro-neurotrophin-4/5, pro-NGF,pro-BDNF, neurotrophin-3, neurotrophin-4/5, NGF, and BDNF. In otherembodiments, the anti-Sortilin antibodies may inhibit interaction (e.g.,binding) between a Sortilin protein of the present disclosure andneurotensin. In other embodiments, the anti-Sortilin antibodies inhibitinteraction (e.g., binding) between a Sortilin protein of the presentdisclosure and p75. In other embodiments, the anti-Sortilin antibodiesmay inhibit interaction (e.g., binding) between a Sortilin protein ofthe present disclosure and a Sort-pro. In other embodiments, theanti-Sortilin antibodies may inhibit interaction (e.g., binding) betweena Sortilin protein of the present disclosure and APP. In otherembodiments, the anti-Sortilin antibodies may inhibit the production ofthe A beta peptide. In other embodiments, the anti-Sortilin antibodiesmay inhibit the transport and secretion of PCSK9. In other embodiments,the anti-Sortilin antibodies may inhibit interaction (e.g., binding)between a Sortilin protein of the present disclosure and LpL. In otherembodiments, the anti-Sortilin antibodies may inhibit interaction (e.g.,binding) between a Sortilin protein of the present disclosure and APOA5.In other embodiments, the anti-Sortilin antibodies may inhibitinteraction (e.g., binding) between a Sortilin protein of the presentdisclosure and APOE. In other embodiments, the anti-Sortilin antibodiesmay inhibit interaction (e.g., binding) between a Sortilin protein ofthe present disclosure and RAP.

In some embodiments, anti-Sortilin antibodies of the present disclosurebind to a Sortilin protein of the present disclosure expressed on thesurface of cell and the naked antibodies inhibit interaction (e.g.,binding) between the Sortilin protein and one or more Sortilin ligands.In some embodiments, anti-Sortilin antibodies of the present disclosurethat bind to a Sortilin protein of the present inhibit interaction(e.g., binding) between the Sortilin protein and one or more Sortilinligands by reducing the effective levels of Sortilin that is availableto interact with these proteins either on the cell surface or inside thecell. In some embodiments, anti-Sortilin antibodies of the presentdisclosure that bind to a Sortilin protein of the present inhibitinteraction (e.g., binding) between the Sortilin protein and one or moreSortilin ligands by inducing degradation of Sortilin.

A. Decreasing interaction between Sortilin and Progranulin

In some embodiments, anti-Sortilin antibodies of the present disclosureincrease Progranulin levels and/or decrease cellular levels of Sortilinwhile blocking (e.g. inhibiting) the interaction (e.g., binding) betweenSortilin and Progranulin. Accordingly, in some embodiments,anti-Sortilin antibodies of the present disclosure block the interaction(e.g., binding) between Sortilin and Progranulin. As used herein, ananti-Sortilin antibody blocks the interaction (e.g., binding) betweenSortilin and Progranulin if it decreases Progranulin binding to Sortilinrelative to a control antibody (e.g. an anti-Sortilin antibody having aheavy chain variable region and a light chain variable regioncorresponding to S-60) at saturating antibody concentrations (e.g., 0.6nM, 0.63 nM, 1.25 nM, 50 nM or 150 nM) in any in vitro assay orcell-based culture assay described herein or known in the art.

Anti-Sortilin antibodies of the present disclosure may decreaseProgranulin binding to Sortilin with a half-maximal effectiveconcentration (EC₅₀) (e.g., when measured in vitro) in the picomolarrange. In certain embodiments, the EC₅₀ of the antibody is less thanabout 2.2 nM. In certain embodiments, the EC₅₀ of the antibody is lessthan about 1.22 nM. In certain embodiments, the EC₅₀ of the antibody isless than about 751 pM. In certain embodiments, the EC₅₀ of the antibodyis about 325 pM to about 751 nM. In certain embodiments, the EC₅₀ of theantibody is about 405 pM to about 751 nM. In certain embodiments, theEC₅₀ of the antibody is about 588 pM to about 751 nM. In certainembodiments, the EC₅₀ of the antibody is less than about 2.2 nM, 2.1 nM,2.0 nM, 1.9 nM, 1.8 nM, 1.7 nM, 1.6 nM, 1.5 nM, 1.4 nM, 1.3 nM, 1.2 nM,1.1 nM, 1.0 nM, 900 pM, 800 pM, 700 pM, 600 pM, 500 pM, 400 pM, 300 pM,200 pM, 100 pM, 50 pM, 40 pM, 30 pM, 20 pM, 10 pM, 1 pM, or 0.5 pM.

In some embodiments, the EC₅₀ of the antibody for decreasing Progranulinbinding to Sortilin is less than about or equal to about 2.2 nM, 2.1 nM,2.0 nM, 1.9 nM, 1.8 nM, 1.7 nM, 1.6 nM, 1.5 nM, 1.4 nM, 1.3 nM, 1.2 nM,1.1 nM, 1.0 nM, 900 pM, 800 pM, 700 pM, 600 pM, 500 pM, 475 pM, 450 pM,425 pM, 400 pM, 375 pM, 350 pM, 325 pM, 300 pM, 275 pM, 250 pM, 225 pM,200 pM, 175 pM, 150 pM, 125 pM, 100 pM, 90 pM, 80 pM, 70 pM, 60 pM, 50pM, 40 pM, 30 pM, 20 pM, 10 pM, 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3pM, 2 pM, 1 pM, or 0.5 pM.

In some embodiments, the EC₅₀ of an anti-Sortilin antibody of thepresent disclosure is about 1.22 nM. In some embodiments, the EC₅₀ of ananti-Sortilin antibody of the present disclosure is about 588 pM. Insome embodiments, the EC₅₀ of an anti-Sortilin antibody of the presentdisclosure is about 405 pM. In some embodiments, the EC₅₀ of ananti-Sortilin antibody of the present disclosure is about 325 pM.

Various methods of measuring antibody EC₅₀ values are known in the art,including, for example, by flow cytometry (See e.g., Example 3). In someembodiments, the EC₅₀ for decreasing Progranulin binding to Sortlin ismeasured in vitro using cells expressing human Sortilin. In someembodiments, the EC₅₀ is measured at a temperature of approximately 4°C. In some embodiments, the EC₅₀ is measured at a temperature ofapproximately 25° C. In some embodiments, the EC₅₀ is measured at atemperature of approximately 35° C. In some embodiments, the EC₅₀ ismeasured at a temperature of approximately 37° C. In some embodiments,the EC₅₀ for decreasing Progranulin binding to Sortlin is determinedusing a monovalent antibody (e.g., a Fab) or a full-length antibody in amonovalent form. In some embodiments, the EC₅₀ is determined usingantibodies containing constant regions that demonstrate enhanced Fcreceptor binding. In some embodiments, the EC₅₀ for decreasingProgranulin binding to Sortlin is determined using antibodies containingconstant regions that demonstrate reduced Fc receptor binding.

In some embodiments, anti-Sortilin antibodies of the present disclosurehave higher potencies in reducing Progranulin binding to Sortlinrelative to a control antibody (e.g. an anti-Sortilin antibody having aheavy chain variable region and a light chain variable regioncorresponding to S-60). In some embodiments, anti-Sortilin antibodies ofthe present disclosure decrease Progranulin binding to Sortlin with alower EC₅₀ (e.g., as measured in vitro) than a control antibody (e.g. ananti-Sortilin antibody having a heavy chain variable region and a lightchain variable region corresponding to S-60). In some embodiments,anti-Sortilin antibodies of the present disclosure decrease Progranulinbinding to Sortlin with an EC₅₀ that is at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, at least about 55%, at least about 60%,at least about 65%, at least about 70%, at least about 75%, at leastabout 80%, at least about 85%, at least about 90%, at least about 95%,or at least about 99% lower than the EC₅₀ of a control antibody (e.g. ananti-Sortilin antibody having a heavy chain variable region and a lightchain variable region corresponding to S-60). In some embodiments,anti-Sortilin antibodies of the present disclosure decrease Progranulinbinding to Sortlin with an EC₅₀ that is at least about 1-fold, at leastabout 1.1-fold, at least about 1.5-fold, at least about 2-fold, at leastabout 3-fold, at least about 4-fold, at least about 5-fold, at leastabout 6-fold, at least about 7-fold, at least about 8-fold, at leastabout 9-fold, at least about 10-fold, at least about 12.5-fold, at leastabout 15-fold, at least about 17.5-fold, at least about 20-fold, atleast about 22.5-fold, at least about 25-fold, at least about 27.5-fold,at least about 30-fold, at least about 50-fold, or at least about100-fold lower than the EC₅₀ of a control antibody (e.g. ananti-Sortilin antibody having a heavy chain variable region and a lightchain variable region corresponding to S-60).

In some embodiments, anti-Sortilin antibodies of the present disclosurehave an EC₅₀ that is at least 1.3-fold lower than control antibody (e.g.an anti-Sortilin antibody having a heavy chain variable region and alight chain variable region corresponding to S-60). In some embodiments,anti-Sortilin antibodies of the present disclosure have an EC₅₀ that isat least 1.8-fold lower than control antibody (e.g. an anti-Sortilinantibody having a heavy chain variable region and a light chain variableregion corresponding to S-60). In some embodiments, anti-Sortilinantibodies of the present disclosure have an EC₅₀ that is at least1.9-fold lower than control antibody (e.g. an anti-Sortilin antibodyhaving a heavy chain variable region and a light chain variable regioncorresponding to S-60). In some embodiments, anti-Sortilin antibodies ofthe present disclosure have an EC₅₀ that is at least 2.3-fold lower thancontrol antibody (e.g. an anti-Sortilin antibody having a heavy chainvariable region and a light chain variable region corresponding toS-60).

Any in vitro cell-based assays or suitable in vivo model describedherein or known in the art may be used to measure inhibition orreduction of interaction (e.g., binding) between Sortilin and one ormore Sortilin ligands. In some embodiments, anti-Sortilin antibodies ofthe present disclosure inhibit or reduce interaction (e.g., binding)between Sortilin and one or more Sortilin ligands by reducing Sortilinexpression (e.g., by reducing cell surface levels of Sortilin). In someembodiments, anti-Sortilin antibodies of the present disclosure inhibitor reduce interaction (e.g., binding) between Sortilin and one or moreSortilin ligands by at least 21%, at least 22%, at least 23%, at least24%, at least 25%, at least 26%, at least 27%, at least 28%, at least29%, at least 30%, at least 31%, at least 32%, at least 33%, at least34%, at least 35%, at least 36%, at least 37%, at least 38%, at least39%, at least 40%, at least 41%, at least 42%, at least 43%, at least44%, at least 45%, at least 46%, at least 47%, at least 48%, at least49%, at least 50%, at least 51%, at least 52%, at least 53%, at least54%, at least 55%, at least 56%, at least 57%, at least 58%, at least59%, at least 60%, at least 61%, at least 62%, at least 63%, at least64%, at least 65%, at least 66%, at least 67%, at least 68%, at least69%, at least 70%, at least 71%, at least 72%, at least 73%, at least74%, at least 75%, at least 76%, at least 77%, at least 78%, at least79%, at least 80%, at least 81%, at least 82%, at least 83%, at least84%, at least 85%, at least 86%, at least 87%, at least 88%, at least89%, at least 90%, at least 91%, at least 92%, at least 93%, at least94%, at least 95%, at least 96%, at least 97%, at least 98%, at least99%, or more at saturating antibody concentrations utilizing any invitro assay or cell-based culture assay described herein or known in theart.

In some embodiments, an anti-Sortilin antibody of the present disclosureblocks Progranulin binding to Sortlin by more than about 90% at 50 nMIgG or by more than about 96% at 150 nM IgG, as measured by flowcytometry. In some embodiments, an anti-Sortilin antibody of the presentdisclosure blocks Progranulin binding to Sortlin by about 90.74% at 50nM IgG, as measured by flow cytometry. In some embodiments, ananti-Sortilin antibody of the present disclosure blocks Progranulinbinding to Sortlin by about 96.5% at 150 nM IgG, as measured by flowcytometry. In some embodiments, an anti-Sortilin antibody of the presentdisclosure blocks Progranulin binding to Sortlin by about 96.9% at 150nM IgG, as measured by flow cytometry.

Decreasing Expression of Pro-Inflammatory Mediators

In some embodiments, anti-Sortilin antibodies of the present disclosuremay decrease the expression of pro-inflammatory mediators after bindingto a Sortilin protein expressed in a cell.

As used herein, pro-inflammatory mediators are proteins involved eitherdirectly or indirectly (e.g., by way of pro-inflammatory signalingpathways) in a mechanism that induces, activates, promotes, or otherwisedecreases an inflammatory response. Any method known in the art foridentifying and characterizing pro-inflammatory mediators may be used.

Examples of pro-inflammatory mediators include, without limitation,cytokines, such as type I and II interferons, IL-6, IL12p70, IL12p40,IL-1β, TNF-α, IL-8, CRP, IL-20 family members, IL-33, LIF, OSM, CNTF,GM-CSF, IL-11, IL-12, IL-17, IL-18, and CRP. Further examples ofpro-inflammatory mediators include, without limitation, chemokines, suchas CXCL1, CCL2, CCL3, CCL4, and CCL5.

In some embodiments, the anti-Sortilin antibodies of the presentdisclosure may decrease functional expression and/or secretion ofpro-inflammatory mediators, IL-6, IL12p70, IL12p40, TNF-α, CXCL1, CCL2,CCL3, CCL4, and CCL5. In certain embodiments, decreased expression ofthe pro-inflammatory mediators occurs in macrophages, dendritic cells,monocytes, osteoclasts, Langerhans cells of skin, Kupffer cells, Tcells, and/or microglial cells. Decreased expression may include,without limitation, a decrease in gene expression, a decrease intranscriptional expression, or a decrease in protein expression. Anymethod known in the art for determining gene, transcript (e.g., mRNA),and/or protein expression may be used. For example, Northern blotanalysis may be used to determine pro-inflammatory mediator geneexpression levels, RT-PCR may be used to determine the level ofpro-inflammatory mediator transcription, and Western blot analysis maybe used to determine pro-inflammatory mediator protein levels.

As used herein, a pro-inflammatory mediator may have decreasedexpression if its expression in one or more cells of a subject treatedwith a Sortilin agent, such as an agonist anti-Sortilin antibody of thepresent disclosure is more than the expression of the samepro-inflammatory mediator expressed in one or more cells of acorresponding subject that is not treated with the agonist anti-Sortilinantibody. In some embodiments, the anti-Sortilin antibody of the presentdisclosure may decrease pro-inflammatory mediator expression in one ormore cells of a subject by at least 10%, at least 15%, at least 20%, atleast 25%, at least 30%, at least 35%, at least 40%, at least 45%, atleast 50%, at least 55%, at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 95%, atleast 100%, at least 110%, at least 115%, at least 120%, at least 125%,at least 130%, at least 135%, at least 140%, at least 145%, at least150%, at least 160%, at least 170%, at least 180%, at least 190%, or atleast 200% for example, as compared to pro-inflammatory mediatorexpression in one or more cells of a corresponding subject that is nottreated with the anti-Sortilin antibody. In other embodiments, theanti-Sortilin antibody may decrease pro-inflammatory mediator expressionin one or more cells of a subject by at least at least 1.5 fold, atleast 1.6 fold, at least 1.7 fold, at least 1.8 fold, at least 1.9 fold,at least 2.0 fold, at least 2.1 fold, at least 2.15 fold, at least 2.2fold, at least 2.25 fold, at least 2.3 fold, at least 2.35 fold, atleast 2.4 fold, at least 2.45 fold, at least 2.5 fold, at least 2.55fold, at least 3.0 fold, at least 3.5 fold, at least 4.0 fold, at least4.5 fold, at least 5.0 fold, at least 5.5 fold, at least 6.0 fold, atleast 6.5 fold, at least 7.0 fold, at least 7.5 fold, at least 8.0 fold,at least 8.5 fold, at least 9.0 fold, at least 9.5 fold, or at least 10fold, for example, as compared to pro-inflammatory mediator expressionin one or more cells of a corresponding subject that is not treated withthe anti-Sortilin antibody.

In some embodiments, an anti-Sortilin antibody according to any of theabove embodiments may incorporate any of the features, singly or incombination, as described in Sections 1-7 below:

(1) Anti-Sortilin Antibody Binding Affinity

In some embodiments of any of the antibodies provided herein, theantibody has a dissociation constant (Kd) of <1 μM, <100 nM, <10 nM, <1nM, <0.1 nM, <0.01 nM, or <0.001 nM (e.g., 10⁻⁸ M or less, e.g., from10⁻⁸ M to 10⁻¹³ M, e.g., from 10⁻⁹ M to 10⁻¹³ M).

Anti-Sortilin antibodies of the present disclosure may have nanomolar oreven picomolar affinities for the target antigen (e.g., human Sortilinor mammalian Sortilin). In certain embodiments, the binding affinity ofan anti-Sortilin antibody of the present disclosure for target antigen(e.g., human Sortilin or mammalian Sortilin) is measured by thedissociation constant, K_(D). Dissociation constants may be determinedthrough any analytical technique, including any biochemical orbiophysical technique such as fluorescent activated cell sorting (FACS),flow cytometry, enzyme-linked immunosorbent assay (ELISA), surfaceplasmon resonance (SPR), BioLayer interferometry (see, e.g., OctetSystem by ForteBio), meso scale discover (see, e.g., MSD-SET),isothermal titration calorimetry (ITC), differential scanningcalorimetry (DSC), circular dichroism (CD), stopped-flow analysis, andcolorimetric or fluorescent protein melting analyses; or a cell bindingassay. In some embodiments, the K_(D) for Sortilin is determined at atemperature of approximately 25° C. In some embodiments, thedissociation constant (K_(D)) may be measured at 4° C. or roomtemperature utilizing, for example, FACS or BioLayer interferometryassay.

In some embodiments, the K_(D) for Sortilin is determined at atemperature of approximately 4° C. In some embodiments, the K_(D) isdetermined using a monovalent antibody (e.g., a Fab) or a full-lengthantibody in a monovalent form. In some embodiments, the K_(D) isdetermined using a bivalent antibody and monomeric recombinant Sortilinprotein.

In certain embodiments, the K_(D) of an anti-Sortilin antibody of thepresent disclosure for human Sortilin, mammalian Sortilin, or both, ismeasured using FACS as described herein (see, e.g., Examples 1 and 4).In certain embodiments, the K_(D) of an anti-Sortilin antibody of thepresent disclosure for human Sortilin, mammalian Sortilin, or both, ismeasured using BioLayer Interferometry as described herein (see, e.g.,Example 4).

In some embodiments, the anti-Sortilin antibody has a dissociationconstant (K_(D)) for human Sortilin that is up to 2.5-fold lower than ananti-Sortilin antibody comprising a heavy chain variable regioncomprising the sequence of SEQ ID NO: 56 and a light chain variableregion comprising the sequence of SEQ ID NO: 79, wherein the K_(D) isdetermined by FACS (see, e.g., Example 1). In some embodiments, theanti-Sortilin antibody has a dissociation constant (K_(D)) for humanSortilin that ranges from about 1.10E-8 M to about 4.68E-10 M whereinthe K_(D) is determined by FACS (see, e.g., Example 1), or about 270 toabout 2910 pM wherein the K_(D) is determined by Bio-layerinterferometry (see, e.g., Example 4).

In certain embodiments, the K_(D) of an anti-Sortilin antibody of thepresent disclosure for human Sortilin, mammalian Sortilin, or both, maybe less than 100 nM, less than 90 nM, less than 80 nM, less than 70 nM,less than 60 nM, less than 50 nM, less than 40 nM, less than 30 nM, lessthan 20 nM, less than 10 nM, less than 9 nM, less than 8 nM, less than 7nM, less than 6 nM, less than 5 nM, less than 4 nM, less than 3 nM, lessthan 2 nM, less than 1 nM, less than 0.5 nM, less than 0.1 nM, less than0.09 nM, less than 0.08 nM, less than 0.07 nM, less than 0.06 nM, lessthan 0.05 nM, less than 0.04 nM, less than 0.03 nM, less than 0.02 nM,less than 0.01 nM, less than 0.009 nM, less than 0.008 nM, less than0.007 nM, less than 0.006 nM, less than 0.005 nM, less than 0.004 nM,less than 0.003 nM, less than 0.002 nM, less than 0.001 nM, or less than0.001 nM.

The dissociation constants (K_(D)) of anti-Sortilin antibodies for humanSortilin, mammalian Sortilin, or both, may be less than 10 nM, less than9.5 nM, less than 9 nM, less than 8.5 nM, less than 8 nM, less than 7.5nM, less than 7 nM, less than 6.9 nM, less than 6.8 nM, less than 6.7nM, less than 6.6 nM, less than 6.5 nM, less than 6.4 nM, less than 6.3nM, less than 6.2 nM, less than 6.1 nM, less than 6 nM, less than 5.5nM, less than 5 nM, less than 4.5 nM, less than 4 nM, less than 3.5 nM,less than 3 nM, less than 2.5 nM, less than 2 nM, less than 1.5 nM, lessthan 1 nM, less than 0.95 nM, less than 0.9 nM, less than 0.89 nM, lessthan 0.88 nM, less than 0.87 nM, less than 0.86 nM, less than 0.85 nM,less than 0.84 nM, less than 0.83 nM, less than 0.82 nM, less than 0.81nM, less than 0.8 nM, less than 0.75 nM, less than 0.7 nM, less than0.65 nM, less than 0.64 nM, less than 0.63 nM, less than 0.62 nM, lessthan 0.61 nM, less than 0.6 nM, less than 0.55 nM, less than 0.5 nM,less than 0.45 nM, less than 0.4 nM, less than 0.35 nM, less than 0.3nM, less than 0.29 nM, less than 0.28 nM, less than 0.27 nM, less than0.26 nM, less than 0.25 nM, less than 0.24 nM, less than 0.23 nM, lessthan 0.22 nM, less than 0.21 nM, less than 0.2 nM, less than 0.15 nM,less than 0.1 nM, less that 0.09 nM, less than 0.08 nM, less than 0.07nM, less than 0.06 nM, less than 0.05 nM, less than 0.04 nM, less than0.03 nM, less than 0.02 nM, less than 0.01 nM, less that 0.009 nM, lessthan 0.008 nM, less than 0.007 nM, less than 0.006 nM, less than 0.005nM, less than 0.004 nM, less than 0.003 nM, less than 0.002 nM, or lessthan 0.001 nM.

In certain embodiments, the dissociation constant (K_(D)) of theantibody for Sortilin is from about 0.560 nM to about 1.63 nM, forexample when the K_(D) is determined by FACS. In certain embodiments,the dissociation constant (K_(D)) of the antibody for Sortilin is fromabout 0.270 nM to about 2.910 nM, for example when the K_(D) isdetermined by BioLayer Interferometry. In some embodiments, the antibodyhas a dissociation constant (K_(D)) for human Sortilin, mouse Sortilin,or both, that ranges from about 0.36 nM to about 0.43 nM, or less than1.02 nM. In some embodiments, the dissociation constant is less than1.02 nM. In some embodiments, an anti-Sortilin antibody of the presentdisclosure has a dissociation constant for human Sortilin of 0.560 nM orless.

In one specific embodiment, an anti-Sortilin antibody of the presentdisclosure has a dissociation constant for human Sortilin of about 0.560nM. In one specific embodiment, an anti-Sortilin antibody of the presentdisclosure has a dissociation constant for human Sortilin of about 0.423nM. In one specific embodiment, an anti-Sortilin antibody of the presentdisclosure has a dissociation constant for human Sortilin of about 0.365nM. In one specific embodiment, an anti-Sortilin antibody of the presentdisclosure has a dissociation constant for human Sortilin of about 0.344nM. In one specific embodiment, an anti-Sortilin antibody of the presentdisclosure has a dissociation constant for human Sortilin of about 0.298nM. In one specific embodiment, an anti-Sortilin antibody of the presentdisclosure has a dissociation constant for human Sortilin of about 0.270nM. In another specific embodiment, an anti-Sortilin antibody of thepresent disclosure has a dissociation constant for human Sortilin ofabout 0.260 nM.

In some embodiments, anti-Sortilin antibodies of the present disclosurehave a lower dissociation constant (K_(D)) for Sortilin than a controlanti-Sortilin antibody (e.g., a control anti-Sortilin antibodycomprising a heavy chain variable region and a light chain variableregion corresponding to S-60. In some embodiments, anti-Sortilinantibodies of the present disclosure have a K_(D) for a target (e.g.,human Sortilin) that is at least about 5%, at least about 10%, at leastabout 15%, at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at leastabout 50%, at least about 55%, at least about 60%, at least about 65%,at least about 70%, at least about 75%, at least about 80%, at leastabout 85%, at least about 90%, at least about 95%, or at least about 99%lower than the K_(D) of a control anti-Sortilin antibody for the target(e.g., a control anti-Sortilin antibody comprising a heavy chainvariable region and a light chain variable region corresponding to S-60.In some embodiments, anti-Sortilin antibodies of the present disclosurehave a K_(D) for a target (e.g., human Sortilin) that is at least about1-fold, at least about 1.1-fold, at least about 1.5-fold, at least about2-fold, at least about 3-fold, at least about 4-fold, at least about5-fold, at least about 6-fold, at least about 7-fold, at least about8-fold, at least about 9-fold, at least about 10-fold, at least about12.5-fold, at least about 15-fold, at least about 17.5-fold, at leastabout 20-fold, at least about 22.5-fold, at least about 25-fold, atleast about 27.5-fold, at least about 30-fold, at least about 50-fold,at least about 100-fold, at least about 200-fold, at least about300-fold, at least about 400-fold, at least about 500-fold, at leastabout 600-fold, at least about 700-fold, at least about 800-fold, atleast about 900-fold, or at least about 1000-fold lower than the K_(D)of a control anti-Sortilin antibody for the target (e.g., a controlanti-Sortilin antibody comprising a heavy chain variable region and alight chain variable region corresponding to S-60.

In some embodiments, anti-Sortilin antibodies of the present disclosurehave a K_(D) for human Sortilin that is at least 100-fold lower than ananti-Sortilin antibody having a heavy chain variable region and a lightchain variable region corresponding to S-60. In some embodiments,anti-Sortilin antibodies of the present disclosure have a K_(D) forhuman Sortilin that is at least 50-fold lower than an anti-Sortilinantibody having a heavy chain variable region and a light chain variableregion corresponding to S-60. In some embodiments, anti-Sortilinantibodies of the present disclosure have a K_(D) for human Sortilinthat is at least 10-fold lower than an anti-Sortilin antibody having aheavy chain variable region and a light chain variable regioncorresponding to S-60. In some embodiments, anti-Sortilin antibodies ofthe present disclosure have a K_(D) for human Sortilin that is at least5-fold lower than an anti-Sortilin antibody having a heavy chainvariable region and a light chain variable region corresponding to S-60.In some embodiments, anti-Sortilin antibodies of the present disclosurehave a K_(D) for human Sortilin that is at least 2-fold lower than ananti-Sortilin antibody having a heavy chain variable region and a lightchain variable region corresponding to S-60.

In a specific embodiment, an anti-Sortilin antibody of the presentdisclosure has a K_(D) for human Sortilin that is about 2.79-fold lowerthan an anti-Sortilin antibody having a heavy chain variable region anda light chain variable region corresponding to S-60. In another specificembodiment, an anti-Sortilin antibody of the present disclosure has aK_(D) for human Sortilin that is about 2.05-fold lower than ananti-Sortilin antibody having a heavy chain variable region and a lightchain variable region corresponding to S-60.

(2) Antibody Fragments

In some embodiments of any of the antibodies provided herein, theantibody antibodies is an antibody fragment. Antibody fragments include,but are not limited to, Fab, Fab′, Fab′-SH, F(ab′)₂, Fv, and scFvfragments, and other fragments described below. For a review of certainantibody fragments, see Hudson et al. Nat. Med. 9:129-134 (2003). For areview of scFv fragments, see, e.g., WO 93/16185; and U.S. Pat. Nos.5,571,894 and 5,587,458. For discussion of Fab and F(ab′)₂ fragmentscomprising salvage receptor binding epitope residues and havingincreased in vivo half-life, see U.S. Pat. No. 5,869,046.

Diabodies are antibody fragments with two antigen-binding sites that maybe bivalent or bispecific. See, for example, EP404097; WO 1993/01161;Hudson et al. Nat. Med. 9:129-134 (2003). Triabodies and tetrabodies arealso described in Hudson et al. Nat. Med. 9:129-134 (2003).Single-domain antibodies are antibody fragments comprising all or aportion of the heavy chain variable domain or all or a portion of thelight chain variable domain of an antibody. In certain embodiments, asingle-domain antibody is a human single-domain antibody (see, e.g.,U.S. Pat. No. 6,248,516).

Antibody fragments can be made by various techniques, including but notlimited to proteolytic digestion of an intact antibody as well asproduction by recombinant host cells (e.g., E. coli or phage), asdescribed herein.

In some embodiments, the antibody fragment is used in combination with asecond Sortilin antibody and/or with one or more antibodies thatspecifically bind a disease-causing protein selected from: amyloid betaor fragments thereof, Tau, IAPP, alpha-synuclein, TDP-43, FUS protein,prion protein, PrPSc, huntingtin, calcitonin, superoxide dismutase,ataxin, Lewy body, atrial natriuretic factor, islet amyloid polypeptide,insulin, apolipoprotein AI, serum amyloid A, medin, prolactin,transthyretin, lysozyme, beta 2 microglobulin, gelsolin,keratoepithelin, cystatin, immunoglobulin light chain AL, S-IBM protein,Repeat-associated non-ATG (RAN) translation products, DiPeptide repeat(DPR) peptides, glycine-alanine (GA) repeat peptides, glycine-proline(GP) repeat peptides, glycine-arginine (GR) repeat peptides,proline-alanine (PA) repeat peptides, proline-arginine (PR) repeatpeptides, and any combination thereof

(3) Chimeric and Humanized Antibodies

In some embodiments of any of the antibodies provided herein, theantibody is a chimeric antibody. Certain chimeric antibodies aredescribed, e.g., in U.S. Pat. No. 4,816,567. In one example, a chimericantibody comprises a non-human variable region (e.g., a variable regionderived from a mouse, rat, hamster, rabbit, or non-human primate, suchas a monkey) and a human constant region. In a further example, achimeric antibody is a “class switched” antibody in which the class orsubclass has been changed from that of the parent antibody. Chimericantibodies include antigen-binding fragments thereof.

In some embodiments of any of the antibodies provided herein, theantibody is a humanized antibody. Typically, a non-human antibody ishumanized to reduce immunogenicity to humans, while retaining thespecificity and affinity of the parental non-human antibody. In certainembodiments, a humanized antibody is substantially non-immunogenic inhumans. In certain embodiments, a humanized antibody has substantiallythe same affinity for a target as an antibody from another species fromwhich the humanized antibody is derived. See, e.g., U.S. Pat. Nos.5,530,101, 5,693,761; 5,693,762; and 5,585,089. In certain embodiments,amino acids of an antibody variable domain that can be modified withoutdiminishing the native affinity of the antigen binding domain whilereducing its immunogenicity are identified. See, e.g., U.S. Pat. Nos.5,766,886 and 5,869,619. Generally, a humanized antibody comprises oneor more variable domains in which HVRs (or portions thereof) are derivedfrom a non-human antibody, and FRs (or portions thereof) are derivedfrom human antibody sequences. A humanized antibody optionally will alsocomprise at least a portion of a human constant region. In someembodiments, some FR residues in a humanized antibody are substitutedwith corresponding residues from a non-human antibody (e.g., theantibody from which the HVR residues are derived), for example, torestore or improve antibody specificity or affinity.

Humanized antibodies and methods of making them are reviewed, forexample, in Almagro et al. Front. Biosci. 13:1619-1633 (2008), and arefurther described, e.g., in U.S. Pat. Nos. 5,821,337, 7,527,791,6,982,321, and 7,087,409. Human framework regions that may be used forhumanization include but are not limited to: framework regions selectedusing the “best-fit” method (see, e.g., Sims et al. J. Immunol. 151:2296(1993)); framework regions derived from the consensus sequence of humanantibodies of a particular subgroup of light or heavy chain variableregions (see, e.g., Carter et al. Proc. Natl. Acad. Sci. USA 89:4285(1992); and Presta et al., J. Immunol. 151:2623 (1993)); human mature(somatically mutated) framework regions or human germline frameworkregions (see, e.g., Almagro and Fransson Front. Biosci. 13:1619-1633(2008)); and framework regions derived from screening FR libraries (see,e.g., Baca et al. J. Biol. Chem. 272:10678-10684 (1997) and Rosok et al.J. Biol. Chem. 271:22611-22618 (1996)).

(4) Human Antibodies

In some embodiments of any of the antibodies provided herein, theantibody is a human antibody. Human antibodies can be produced usingvarious techniques known in the art. Human antibodies are describedgenerally in van Dijk et al. Curr. Opin. Pharmacol. 5:368-74 (2001) andLonberg Curr. Opin. Immunol. 20:450-459 (2008).

Human antibodies may be prepared by administering an immunogen to atransgenic animal that has been modified to produce intact humanantibodies or intact antibodies with human variable regions in responseto antigenic challenge. One can engineer mouse strains deficient inmouse antibody production with large fragments of the human Ig loci inanticipation that such mice would produce human antibodies in theabsence of mouse antibodies. Large human Ig fragments can preserve thelarge variable gene diversity as well as the proper regulation ofantibody production and expression. By exploiting the mouse machineryfor antibody diversification and selection and the lack of immunologicaltolerance to human proteins, the reproduced human antibody repertoire inthese mouse strains can yield high affinity fully human antibodiesagainst any antigen of interest, including human antigens. Using thehybridoma technology, antigen-specific human MAbs with the desiredspecificity can be produced and selected. Certain exemplary methods aredescribed in U.S. Pat. No. 5,545,807, EP 546073, and EP 546073. Seealso, for example, U.S. Pat. Nos. 6,075,181 and 6,150,584 describingXENOMOUSE™ technology; U.S. Pat. No. 5,770,429 describing HUMAB®technology; U.S. Pat. No. 7,041,870 describing K-M MOUSE® technology,and U.S. Patent Application Publication No. US 2007/0061900, describingVELOCIMOUSE® technology. Human variable regions from intact antibodiesgenerated by such animals may be further modified, e.g., by combiningwith a different human constant region.

Human antibodies can also be made by hybridoma-based methods. Humanmyeloma and mouse-human heteromyeloma cell lines for the production ofhuman monoclonal antibodies have been described. (See, e.g., Kozbori J.Immunol. 133:3001 (1984) and Boerner et al. J. Immunol. 147:86 (1991)).Human antibodies generated via human B-cell hybridoma technology arealso described in Li et al. Proc. Natl. Acad. Sci. USA, 103:3557-3562(2006). Additional methods include those described, for example, in U.S.Pat. No. 7,189,826 (describing production of monoclonal human IgMantibodies from hybridoma cell lines). Human hybridoma technology(Trioma technology) is also described in Vollmers et al. Histology andHistopathology 20(3):927-937 (2005) and Vollmers et al. Methods andFindings in Experimental and Clinical Pharmacology 27(3):185-91 (2005).Human antibodies may also be generated by isolating Fv clone variabledomain sequences selected from human-derived phage display libraries.Such variable domain sequences may then be combined with a desired humanconstant domain. Techniques for selecting human antibodies from antibodylibraries are described below.

In some embodiments of any of the antibodies provided herein, theantibody is a human antibody isolated by in vitro methods and/orscreening combinatorial libraries for antibodies with the desiredactivity or activities. Suitable examples include but are not limited tophage display (CAT, Morphosys, Dyax, Biosite/Medarex, Xoma, Symphogen,Alexion (formerly Proliferon), Affimed) ribosome display (CAT),yeast-based platforms (Adimab), and the like. In certain phage displaymethods, repertoires of VH and VL genes are separately cloned bypolymerase chain reaction (PCR) and recombined randomly in phagelibraries, which can then be screened for antigen-binding phage asdescribed in Winter et al. Ann. Rev. Immunol. 12: 433-455 (1994). Forexample, a variety of methods are known in the art for generating phagedisplay libraries and screening such libraries for antibodies possessingthe desired binding characteristics. See also Sidhu et al. J. Mol. Biol.338(2): 299-310, 2004; Lee et al. J. Mol. Biol. 340(5): 1073-1093, 2004;Fellouse Proc. Natl. Acad. Sci. USA 101(34):12467-12472 (2004); and Leeet al. J. Immunol. Methods 284(-2):119-132 (2004). Phage typicallydisplay antibody fragments, either as single-chain Fv (scFv) fragmentsor as Fab fragments. Libraries from immunized sources providehigh-affinity antibodies to the immunogen without the requirement ofconstructing hybridomas. Alternatively, the naive repertoire can becloned (e.g., from human) to provide a single source of antibodies to awide range of non-self and also self-antigens without any immunizationas described by Griffiths et al. EMBO J. 12: 725-734 (1993). Finally,naive libraries can also be made synthetically by cloning unrearrangedV-gene segments from stem cells, and using PCR primers comprising randomsequence to encode the highly variable HVR3 regions and to accomplishrearrangement in vitro, as described by Hoogenboom et al. J. Mol. Biol.,227: 381-388, 1992. Patent publications describing human antibody phagelibraries include, for example: U.S. Pat. No. 5,750,373, and US PatentPublication Nos. 2007/0292936 and 2009/0002360. Antibodies isolated fromhuman antibody libraries are considered human antibodies or humanantibody fragments herein.

(5) Constant Regions Including Fc Regions

In some embodiments of any of the antibodies provided herein, theantibody comprises an Fc. In some embodiments, the Fc is a human IgG1,IgG2, IgG3, and/or IgG4 isotype. In some embodiments, the antibody is ofthe IgG class, the IgM class, or the IgA class.

In certain embodiments of any of the antibodies provided herein, theantibody has an IgG2 isotype. In some embodiments, the antibody containsa human IgG2 constant region. In some embodiments, the human IgG2constant region includes an Fc region. In some embodiments, the antibodyinduces the one or more Sortilin activities or independently of bindingto an Fc receptor. In some embodiments, the antibody binds an inhibitoryFc receptor. In certain embodiments, the inhibitory Fc receptor isinhibitory Fc-gamma receptor IIB (FcγIIB).

In certain embodiments of any of the antibodies provided herein, theantibody has an IgG1 isotype. In some embodiments, the antibody containsa mouse IgG1 constant region. In some embodiments, the antibody containsa human IgG1 constant region. In some embodiments, the human IgG1constant region includes an Fc region. In some embodiments, the antibodybinds an inhibitory Fc receptor. In certain embodiments, the inhibitoryFc receptor is inhibitory Fc-gamma receptor IIB (FcγIIB).

In certain embodiments of any of the antibodies provided herein, theantibody has an IgG4 isotype. In some embodiments, the antibody containsa human IgG4 constant region. In some embodiments, the human IgG4constant region includes an Fc region. In some embodiments, the antibodybinds an inhibitory Fc receptor. In certain embodiments, the inhibitoryFc receptor is inhibitory Fc-gamma receptor IIB (FcγIIB).

In certain embodiments of any of the antibodies provided herein, theantibody has a hybrid IgG2/4 isotype. In some embodiments, the antibodyincludes an amino acid sequence comprising amino acids 118 to 260according to EU numbering of human IgG2 and amino acids 261-447according to EU numbering of human IgG4 (WO 1997/11971; WO 2007/106585).

In some embodiments, the Fc region increases clustering withoutactivating complement as compared to a corresponding antibody comprisingan Fc region that does not comprise the amino acid substitutions. Insome embodiments, the antibody induces one or more activities of atarget specifically bound by the antibody. In some embodiments, theantibody binds to Sortilin.

It may also be desirable to modify an anti-Sortilin antibody of thepresent disclosure to modify effector function and/or to increase serumhalf-life of the antibody. For example, the Fc receptor binding site onthe constant region may be modified or mutated to remove or reducebinding affinity to certain Fc receptors, such as FcγRI, FcγRII, and/orFcγRIII to reduce Antibody-dependent cell-mediated cytotoxicity. In someembodiments, the effector function is impaired by removingN-glycosylation of the Fc region (e.g., in the CH2 domain of IgG) of theantibody. In some embodiments, the effector function is impaired bymodifying regions such as 233-236, 297, and/or 327-331 of human IgG asdescribed in WO 99/58572 and Armour et al. Molecular Immunology 40:585-593 (2003); Reddy et al. J. Immunology 164:1925-1933 (2000). Inother embodiments, it may also be desirable to modify an anti-Sortilinantibody of the present disclosure to modify effector function toincrease finding selectivity toward the ITIM-containing FcgRIIb (CD32b)to increase clustering of Sortilin antibodies on adjacent cells withoutactivating humoral responses including Antibody-dependent cell-mediatedcytotoxicity and antibody-dependent cellular phagocytosis.

To increase the serum half-life of the antibody, one may incorporate asalvage receptor binding epitope into the antibody (especially anantibody fragment) as described in U.S. Pat. No. 5,739,277, for example.As used herein, the term “salvage receptor binding epitope” refers to anepitope of the Fc region of an IgG molecule (e.g., IgG₁, IgG₂, IgG₃, orIgG₄) that is responsible for increasing the in vivo serum half-life ofthe IgG molecule. Other amino acid sequence modifications.

(6) Multispecific Antibodies

Multispecific are antibodies that have binding specificities for atleast two different epitopes, including those on the same or anotherpolypeptide (e.g., one or more Sortilin polypeptides of the presentdisclosure). In some embodiments, the multispecific antibody can be abispecific antibody. In some embodiments, the multispecific antibody canbe a trispecific antibody. In some embodiments, the multispecificantibody can be a tetraspecific antibody. Such antibodies can be derivedfrom full-length antibodies or antibody fragments (e.g., F(ab′)₂bispecific antibodies). In some embodiments, the multispecific antibodycomprises a first antigen binding region which binds to first site onSortilin and comprises a second antigen binding region which binds to asecond site on Sortilin. In some embodiment, the multispecificantibodies comprises a first antigen binding region which binds toSortilin and a second antigen binding region that binds to a secondpolypeptide.

Provided herein are multispecific antibodies comprises a first antigenbinding region, wherein the first antigen binding region comprises thesix HVRs of an antibody described herein, which binds to Sortilin and asecond antigen binding region that binds to a second polypeptide. Insome embodiments, the first antigen binding region comprises the V_(H)or V_(L) of an antibody described herein.

In some embodiments of any of the multispecific antibodies, the secondpolypeptide is a) an antigen facilitating transport across theblood-brain-barrier; (b) an antigen facilitating transport across theblood-brain-barrier selected from transferrin receptor (TR), insulinreceptor (HIR), insulin-like growth factor receptor (IGFR), low-densitylipoprotein receptor related proteins 1 and 2 (LPR-1 and 2), diphtheriatoxin receptor, CRM197, a llama single domain antibody, TMEM 30(A), aprotein transduction domain, TAT, Syn-B, penetratin, a poly-argininepeptide, an angiopep peptide, and ANG1005; (c) a disease-causing proteinselected from amyloid beta, oligomeric amyloid beta, amyloid betaplaques, amyloid precursor protein or fragments thereof, Tau, IAPP,alpha-synuclein, TDP-43, FUS protein, C9orf72 (chromosome 9 open readingframe 72), c9RAN protein, prion protein, PrPSc, huntingtin, calcitonin,superoxide dismutase, ataxin, ataxin 1, ataxin 2, ataxin 3, ataxin 7,ataxin 8, ataxin 10, Lewy body, atrial natriuretic factor, islet amyloidpolypeptide, insulin, apolipoprotein AI, serum amyloid A, medin,prolactin, transthyretin, lysozyme, beta 2 microglobulin, gelsolin,keratoepithelin, cystatin, immunoglobulin light chain AL, S-IBM protein,Repeat-associated non-ATG (RAN) translation products, DiPeptide repeat(DPR) peptides, glycine-alanine (GA) repeat peptides, glycine-proline(GP) repeat peptides, glycine-arginine (GR) repeat peptides,proline-alanine (PA) repeat peptides, ubiquitin, and proline-arginine(PR) repeat peptides; (d) ligands and/or proteins expressed on immunecells, wherein the ligands and/or proteins selected from CD40, OX40,ICOS, CD28, CD137/4-1BB, CD27, GITR, PD-L1, CTLA-4, PD-L2, PD-1, B7-H3,B7-H4, HVEM, BTLA, KIR, GALS, TIM3, A2AR, LAG-3, and phosphatidylserine;and/or (e) a protein, lipid, polysaccharide, or glycolipid expressed onone or more tumor cells and any combination thereof.

Numerous antigens are known in the art that facilitate transport acrossthe blood-brain barrier (see, e.g., Gabathuler R. Neurobiol. Dis.37:48-57 (2010)). Such second antigens include, without limitation,transferrin receptor (TR), insulin receptor (HIR), Insulin-like growthfactor receptor (IGFR), low-density lipoprotein receptor relatedproteins 1 and 2 (LPR-1 and 2), diphtheria toxin receptor, includingCRM197 (a non-toxic mutant of diphtheria toxin), llama single domainantibodies such as TMEM 30(A) (Flippase), protein transduction domainssuch as TAT, Syn-B, or penetratin, poly-arginine or generally positivelycharged peptides, Angiopep peptides such as ANG1005 (see, e.g.,Gabathuler, 2010), and other cell surface proteins that are enriched onblood-brain barrier endothelial cells (see, e.g., Daneman et al. PLoSOne 5(10):e13741 (2010)).

The multivalent antibodies may recognize the Sortilin antigen as well aswithout limitation additional antigens Aβ peptide, antigen or anα-synuclein protein antigen or, Tau protein antigen or, TDP-43 proteinantigen or, prion protein antigen or, huntingtin protein antigen, orRAN, translation Products antigen, including the DiPeptide Repeats,(DPRs peptides) composed of glycine-alanine (GA), glycine-proline (GP),glycine-arginine (GR), proline-alanine (PA), or proline-arginine (PR),Insulin receptor, insulin like growth factor receptor. Transferrinreceptor or any other antigen that facilitate antibody transfer acrossthe blood brain barrier. In some embodiments, the second polypeptide istransferrin. In some embodiments, the second polypeptide is Tau. In someembodiments, the second polypeptide is Aβ. In some embodiments, thesecond polypeptide is TREM2. In some embodiments, the second polypeptideis α-synuclein.

The multivalent antibody contains at least one polypeptide chain (andpreferably two polypeptide chains), wherein the polypeptide chain orchains comprise two or more variable domains. For instance, thepolypeptide chain or chains may comprise VD1-(X1)_(n)-VD2-(X2)_(n)-Fc,wherein VD1 is a first variable domain, VD2 is a second variable domain,Fc is one polypeptide chain of an Fc region, X1 and X2 represent anamino acid or polypeptide, and n is 0 or 1. Similarly, the polypeptidechain or chains may comprise V_(H)-C_(H)1-flexiblelinker-V_(H)-C_(H)1-Fc region chain; or V_(H)-C_(H)1-V_(H)-C_(H)1-Fcregion chain. The multivalent antibody herein preferably furthercomprises at least two (and preferably four) light chain variable domainpolypeptides. The multivalent antibody herein may, for instance,comprise from about two to about eight light chain variable domainpolypeptides. The light chain variable domain polypeptides contemplatedhere comprise a light chain variable domain and, optionally, furthercomprise a CL domain.

Techniques for making multispecific antibodies include, but are notlimited to, recombinant co-expression of two immunoglobulin heavychain-light chain pairs having different specificities (see Milstein andCuello Nature 305: 537 (1983), WO 93/08829, and Traunecker et al. EMBOJ. 10:3655 (1991)), and “knob-in-hole” engineering (see, e.g., U.S. Pat.No. 5,731,168). See also WO 2013/026833 (CrossMab). Multi-specificantibodies may also be made by engineering electrostatic steeringeffects for making antibody Fc-heterodimeric molecules (WO2009/089004A1); cross-linking two or more antibodies (see, e.g., U.S.Pat. No. 4,676,980); using leucine; using “diabody” technology formaking bispecific antibody fragments (see, e.g., Hollinger et al. Proc.Natl. Acad. Sci. USA 90:6444-6448 (1993)); and using single-chain Fv(scFv) dimers (see, e.g., Gruber et al. J. Immunol. 152:5368 (1994));and preparing trispecific antibodies as described, e.g., in Tutt et al.J. Immunol. 147: 60 (1991).

Engineered antibodies with three or more functional antigen bindingsites, including “Octopus antibodies,” are also included herein (see,e.g., US 2006/0025576). The antibody herein also includes a “Dual ActingFAb” or “DAF” comprising an antigen binding site that binds to multipleSortilin (see, US 2008/0069820, for example).

(7) Antibodies with Improved Stability

Amino acid sequence modifications of anti-Sortilin antibodies of thepresent disclosure, or antibody fragments thereof to improve stabilityduring manufacturing, storage, and in vivo administration, are alsocontemplated. For example, it may be desirable to reduce degradation ofthe antibodies or antibody fragments of the present disclosure throughmultiple pathways, including without limitation, oxidation anddeamidation. Amino acid sequence variants of the antibodies or antibodyfragments are prepared by introducing appropriate nucleotide changesinto the nucleic acid encoding the antibodies or antibody fragments, orby peptide synthesis. Such modifications include, for example, deletionsfrom, and/or insertions into and/or substitutions of, residues withinthe amino acid sequences of the antibody. Any combination of deletion,insertion, and substitution can be made to arrive at the finalconstruct, provided that the final construct possesses the desiredcharacteristics (i.e., reduced susceptibility to degradation).

In some embodiments, the asparagine (N33) site in the HVR-L1 region ofan anti-Sortilin antibody of the present disclosure may be susceptibleto degradation by means of deamidation. In certain embodiments, theasparagine (N33) site in the HVR-L1 region of S-60-15 (SEQ ID NO:8) maybe susceptible to deamidation. Upon deamidation, the asparagine (N33)site in the HVR-L1 region of S-60-15 results in an Asn to Asp/IsoAspchange. In certain embodiments, the asparagine (N33) site in the HVR-L1region of S-60-15 may be substituted to prevent or reduce deamidation.Non-limiting exemplary amino acid sequence variants of S-60-15 havingamino acid substitutions in the asparagine (N33) site of the HVR-L1region include S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3 [N33G],S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7 [N33K],S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E], S-60-15.11[N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V],S-60-15.15 [N33A], S-60-15.16 [N33M], or S-60-15.17 [N33L].

(8) Antibody Variants

In some embodiments of any of the antibodies provided herein, amino acidsequence variants of the antibodies are contemplated. For example, itmay be desirable to improve the binding affinity and/or other biologicalproperties of the antibody.

(i) Substitution, Insertion, and Deletion Variants

In some embodiments of any of the antibodies provided herein, antibodyvariants having one or more amino acid substitutions are provided. Aminoacid sequence variants of an antibody may be prepared by introducingappropriate modifications into the nucleotide sequence encoding theantibody, or by peptide synthesis. Such modifications include, forexample, deletions from, and/or insertions into and/or substitutions ofresidues within the amino acid sequences of the antibody.

TABLE 37 Amino Acid Substitutions Original Exemplary Preferred ResidueSubstitutions Substitutions Ala (A) Val; Leu; Ile Val Arg (R) Lys; Gln;Asn Lys Asn (N) Gln; His; Asp, Lys; Arg Gln Asp (D) Glu; Asn Glu Cys (C)Ser; Ala Ser Gln (Q) Asn; Glu Asn Glu (E) Asp; Gln Asp Gly (G) Ala AlaHis (H) Asn; Gln; Lys; Arg Arg Ile (I) Leu; Val; Met; Ala; Phe;Norleucine Leu Leu (L) Norleucine; Ile; Val; Met; Ala; Phe Ile Lys (K)Arg; Gln; Asn Arg Met (M) Leu; Phe; Ile Leu Phe (F) Leu; Val; Ile; Ala;Tyr Tyr Pro (P) Ala Ala Ser (S) Thr Thr Thr (T) Ser Ser Trp (W) Tyr; PheTyr Tyr (Y) Trp; Phe; Thr; Ser Phe Val (V) Ile; Leu; Met; Phe; Ala;Norleucine Leu

Substantial modifications in the biological properties of the antibodyare accomplished by selecting substitutions that differ significantly intheir effect on maintaining (a) the structure of the polypeptidebackbone in the area of the substitution, for example, as a sheet orhelical conformation, (b) the charge or hydrophobicity of the moleculeat the target site, or (c) the bulk of the side chain. Naturallyoccurring residues are divided into groups based on common side-chainproperties:

-   -   (1) hydrophobic: Norleucine, Met, Ala, Val, Leu, Ile;    -   (2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gln;    -   (3) acidic: Asp, Glu;    -   (4) basic: His, Lys, Arg;    -   (5) residues that influence chain orientation: Gly, Pro; and    -   (6) aromatic: Trp, Tyr, Phe.

For example, non-conservative substitutions can involve the exchange ofa member of one of these classes for a member from another class. Suchsubstituted residues can be introduced, for example, into regions of ahuman antibody that are homologous with non-human antibodies, or intothe non-homologous regions of the molecule.

In making changes to the polypeptide or antibody described herein,according to certain embodiments, the hydropathic index of amino acidscan be considered. Each amino acid has been assigned a hydropathic indexon the basis of its hydrophobicity and charge characteristics. They are:isoleucine (+4.5); valine (+4.2); leucine (+3.8); phenylalanine (+2.8);cysteine/cystine (+2.5); methionine (+1.9); alanine (+1.8); glycine(−0.4); threonine (−0.7); serine (−0.8); tryptophan (−0.9); tyrosine(−1.3); proline (−1.6); histidine (−3.2); glutamate (−3.5); glutamine(−3.5); aspartate (−3.5); asparagine (−3.5); lysine (−3.9); and arginine(−4.5).

The importance of the hydropathic amino acid index in conferringinteractive biological function on a protein is understood in the art.Kyte et al. J. Mol. Biol., 157:105-131 (1982). It is known that certainamino acids can be substituted for other amino acids having a similarhydropathic index or score and still retain a similar biologicalactivity. In making changes based upon the hydropathic index, in certainembodiments, the substitution of amino acids whose hydropathic indicesare within ±2 is included. In certain embodiments, those which arewithin ±1 are included, and in certain embodiments, those within ±0.5are included.

It is also understood in the art that the substitution of like aminoacids can be made effectively on the basis of hydrophilicity,particularly where the biologically functional protein or peptidethereby created is intended for use in immunological embodiments, as inthe present case. In certain embodiments, the greatest local averagehydrophilicity of a protein, as governed by the hydrophilicity of itsadjacent amino acids, correlates with its immunogenicity andantigenicity, i.e., with a biological property of the protein.

The following hydrophilicity values have been assigned to these aminoacid residues: arginine (+3.0); lysine (+3.0±1); aspartate (+3.0±1);glutamate (+3.0±1); serine (+0.3); asparagine (+0.2); glutamine (+0.2);glycine (0); threonine (−0.4); proline (−0.5±1); alanine (−0.5);histidine (−0.5); cysteine (−1.0); methionine (−1.3); valine (−1.5);leucine (−1.8); isoleucine (−1.8); tyrosine (−2.3); phenylalanine (−2.5)and tryptophan (−3.4). In making changes based upon similarhydrophilicity values, in certain embodiments, the substitution of aminoacids whose hydrophilicity values are within ±2 is included, in certainembodiments, those which are within ±1 are included, and in certainembodiments, those within ±0.5 are included. One can also identifyepitopes from primary amino acid sequences on the basis ofhydrophilicity. These regions are also referred to as “epitopic coreregions”.

In certain embodiments, substitutions, insertions, or deletions mayoccur within one or more HVRs so long as such alterations do notsubstantially reduce the ability of the antibody to bind antigen. Forexample, conservative alterations (e.g., conservative substitutions asprovided herein) that do not substantially reduce binding affinity maybe made in HVRs. Such alterations may, for example, be outside ofantigen contacting residues in the HVRs. In certain embodiments of thevariant VH and VL sequences provided above, each HVR either isunaltered, or contains no more than one, two or three amino acidsubstitutions.

Amino acid sequence insertions include amino- and/or carboxyl-terminalfusions ranging in length from one residue to polypeptides comprising ahundred or more residues, as well as intrasequence insertions of singleor multiple amino acid residues. Examples of terminal insertions includean antibody with an N-terminal methionyl residue. Other insertionalvariants of the antibody molecule include the fusion to the N- orC-terminus of the antibody to an enzyme (e.g., for ADEPT) or apolypeptide which increases the serum half-life of the antibody.

Any cysteine residue not involved in maintaining the proper conformationof the antibody also may be substituted, generally with serine, toimprove the oxidative stability of the molecule and prevent aberrantcrosslinking. Conversely, cysteine bond(s) may be added to the antibodyto improve its stability (particularly where the antibody is an antibodyfragment, such as an Fv fragment).

(ii) Glycosylation Variants

In some embodiments of any of the antibodies provided herein, theantibody is altered to increase or decrease the extent to which theantibody is glycosylated. Addition or deletion of glycosylation sites toan antibody may be conveniently accomplished by altering the amino acidsequence such that one or more glycosylation sites is created orremoved.

Glycosylation of antibodies is typically either N-linked or O-linked.N-linked refers to the attachment of the carbohydrate moiety to the sidechain of an asparagine residue. The tripeptide sequencesasparagine-X-serine and asparagine-X-threonine, where X is any aminoacid except proline, are the recognition sequences for enzymaticattachment of the carbohydrate moiety to the asparagine side chain.Thus, the presence of either of these tripeptide sequences in apolypeptide creates a potential glycosylation site. O-linkedglycosylation refers to the attachment of one of the sugarsN-aceylgalactosamine, galactose, or xylose to a hydroxyamino acid, mostcommonly serine or threonine, although 5-hydroxyproline or5-hydroxylysine may also be used.

Addition of glycosylation sites to the antibody is convenientlyaccomplished by altering the amino acid sequence such that it containsone or more of the above-described tripeptide sequences (for N-linkedglycosylation sites). The alteration may also be made by the additionof, or substitution by, one or more serine or threonine residues to thesequence of the original antibody (for O-linked glycosylation sites).

Where the antibody comprises an Fc region, the carbohydrate attachedthereto may be altered. Native antibodies produced by mammalian cellstypically comprise a branched, biantennary oligosaccharide that isgenerally attached by an N-linkage to Asn297 according to Kabatnumbering of the CH2 domain of the Fc region. The oligosaccharide mayinclude various carbohydrates, for example, mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as a fucoseattached to a GlcNAc in the “stem” of the biantennary oligosaccharidestructure. In some embodiments, modifications of the oligosaccharide inan antibody of the invention may be made in order to create antibodyvariants with certain improved properties.

In one embodiment, antibody variants are provided having a carbohydratestructure that lacks fucose attached (directly or indirectly) to an Fcregion. See, e.g., US Patent Publication Nos. 2003/0157108 and2004/0093621. Examples of publications related to “defucosylated” or“fucose-deficient” antibody variants include: US 2003/0157108; US2003/0115614; US 2002/0164328; US 2004/0093621; US 2004/0132140; US2004/0110704; US 2004/0110282; US 2004/0109865; Okazaki et al. J. Mol.Biol. 336:1239-1249 (2004); Yamane-Ohnuki et al. Biotech. Bioeng. 87:614(2004). Examples of cell lines capable of producing defucosylatedantibodies include Led 3 CHO cells deficient in protein fucosylation(Ripka et al. Arch. Biochem. Biophys. 249:533-545 (1986); US2003/0157108), and knockout cell lines, such asalpha-1,6-fucosyltransferase gene, FUT8, knockout CHO cells (see, e.g.,Yamane-Ohnuki et al. Biotech. Bioeng. 87: 614 (2004) and Kanda et al.Biotechnol. Bioeng. 94(4):680-688 (2006)).

(iii) Modified Constant Regions

In some embodiments of any of the antibodies provided herein, theantibody Fc is an antibody Fc isotypes and/or modifications. In someembodiments, the antibody Fc isotype and/or modification is capable ofbinding to Fc gamma receptor.

In some embodiments of any of the antibodies provided herein, themodified antibody Fc is an IgG1 modified Fc. In some embodiments, theIgG1 modified Fc comprises one or more modifications. For example, insome embodiments, the IgG1 modified Fc comprises one or more amino acidsubstitutions (e.g., relative to a wild-type Fc region of the sameisotype). In some embodiments, the one or more amino acid substitutionsare selected from N297A (Bolt S et al. (1993) Eur J Immunol 23:403-411),D265A (Shields et al. (2001) R. J. Biol. Chem. 276, 6591-6604), L234A,L235A (Hutchins et al. (1995) Proc Natl Acad Sci USA, 92:11980-11984;Alegre et al., (1994) Transplantation 57:1537-1543. 31; Xu et al.,(2000) Cell Immunol, 200:16-26), G237A (Alegre et al. (1994)Transplantation 57:1537-1543. 31; Xu et al. (2000) Cell Immunol,200:16-26), C226S, C229S, E233P, L234V, L234F, L235E (McEarchern et al.,(2007) Blood, 109:1185-1192), P331S (Sazinsky et al., (2008) Proc NatlAcad Sci USA 2008, 105:20167-20172), S267E, L328F, A330L, M252Y, S254T,and/or T256E, where the amino acid position is according to the EUnumbering convention. In some embodiments of any of the antibodiesprovided herein, the antibody is an IgG1 isotype and the Fc regioncomprises amino acid substitutions at positions L234A, L235A, and P331S,wherein the numbering of the residue position is according to EUnumbering.

In some embodiments of any of the IgG1 modified Fc, the Fc comprisesN297A mutation according to EU numbering. In some embodiments of any ofthe IgG1 modified Fc, the Fc comprises D265A and N297A mutationsaccording to EU numbering. In some embodiments of any of the IgG1modified Fc, the Fc comprises D270A mutations according to EU numbering.In some embodiments, the IgG1 modified Fc comprises L234A and L235Amutations according to EU numbering. In some embodiments of any of theIgG1 modified Fc, the Fc comprises L234A and G237A mutations accordingto EU numbering. In some embodiments of any of the IgG1 modified Fc, theFc comprises L234A, L235A and G237A mutations according to EU numbering.In some embodiments of any of the IgG1 modified Fc, the Fc comprises oneor more (including all) of P238D, L328E, E233, G237D, H268D, P271G andA330R mutations according to EU numbering. In some embodiments of any ofthe IgG1 modified Fc, the Fc comprises one or more of S267E/L328Fmutations according to EU numbering. In some embodiments of any of theIgG1 modified Fc, the Fc comprises P238D, L328E, E233D, G237D, H268D,P271G and A330R mutations according to EU numbering. In some embodimentsof any of the IgG1 modified Fc, the Fc comprises P238D, L328E, G237D,H268D, P271G and A330R mutations according to EU numbering. In someembodiments of any of the IgG1 modified Fc, the Fc comprises P238D,S267E, L328E, E233D, G237D, H268D, P271G and A330R mutations accordingto EU numbering. In some embodiments of any of the IgG1 modified Fc, theFc comprises P238D, S267E, L328E, G237D, H268D, P271G and A330Rmutations according to EU numbering. In some embodiments of any of theIgG1 modified Fc, the Fc comprises C226S, C229S, E233P, L234V, and L235Amutations according to EU numbering. In some embodiments of any of theIgG1 modified Fc, the Fc comprises L234F, L235E, and P331S mutationsaccording to EU numbering. In some embodiments of any of the IgG1modified Fc, the Fc comprises S267E and L328F mutations according to EUnumbering. In some embodiments of any of the IgG1 modified Fc, the Fccomprises S267E mutations according to EU numbering. In some embodimentsof any of the IgG1 modified Fc, the Fc comprises a substitute of theconstant heavy 1 (CH1) and hinge region of IgG1 with CH1 and hingeregion of IgG2 (amino acids 118-230 of IgG2 according to EU numbering)with a Kappa light chain.

In some embodiments of any of the IgG1 modified Fc, the Fc includes twoor more amino acid substitutions that increase antibody clusteringwithout activating complement as compared to a corresponding antibodyhaving an Fc region that does not include the two or more amino acidsubstitutions. Accordingly, in some embodiments of any of the IgG1modified Fc, the IgG1 modified Fc is an antibody comprising an Fcregion, where the antibody comprises an amino acid substitution atposition E430G and one or more amino acid substitutions in the Fc regionat a residue position selected from: L234F, L235A, L235E, S267E, K322A,L328F, A330S, P331S, and any combination thereof according to EUnumbering. In some embodiments, the IgG1 modified Fc comprises an aminoacid substitution at positions E430G, L243A, L235A, and P331S accordingto EU numbering. In some embodiments, the IgG1 modified Fc comprises anamino acid substitution at positions E430G and P331S according to EUnumbering. In some embodiments, the IgG1 modified Fc comprises an aminoacid substitution at positions E430G and K322A according to EUnumbering. In some embodiments, the IgG1 modified Fc comprises an aminoacid substitution at positions E430G, A330S, and P331S according to EUnumbering. In some embodiments, the IgG1 modified Fc comprises an aminoacid substitution at positions E430G, K322A, A330S, and P331S accordingto EU numbering. In some embodiments, the IgG1 modified Fc comprises anamino acid substitution at positions E430G, K322A, and A330S accordingto EU numbering. In some embodiments, the IgG1 modified Fc comprises anamino acid substitution at positions E430G, K322A, and P331S accordingto EU numbering.

In some embodiments of any of the IgG1 modified Fc, the IgG1 modified Fcmay further comprise herein may be combined with an A330L mutation(Lazar et al. Proc Natl Acad Sci USA, 103:4005-4010 (2006)), or one ormore of L234F, L235E, and/or P331S mutations (Sazinsky et al. Proc NatlAcad Sci USA, 105:20167-20172 (2008)), according to the EU numberingconvention, to eliminate complement activation. In some embodiments ofany of the IgG1 modified Fc, the IgG1 modified Fc may further compriseone or more of A330L, A330S, L234F, L235E, and/or P331S according to EUnumbering. In some embodiments of any of the IgG1 modified Fc, the IgG1modified Fc may further comprise one or more mutations to enhance theantibody half-life in human serum (e.g., one or more (including all) ofM252Y, S254T, and T256E mutations according to the EU numberingconvention). In some embodiments of any of the IgG1 modified Fc, theIgG1 modified Fc may further comprise one or more of E430G, E430S,E430F, E430T, E345K, E345Q, E345R, E345Y, S440Y, and/or S440W accordingto EU numbering.

Other aspects of the present disclosure relate to antibodies havingmodified constant regions (i.e., Fc regions). An antibody dependent onbinding to FcgR receptor to activate targeted receptors may lose itsagonist activity if engineered to eliminate FcgR binding (see, e.g.,Wilson et al. Cancer Cell 19:101-113 (2011); Armour at al. Immunology40:585-593 (2003); and White et al. Cancer Cell 27:138-148 (2015)). Assuch, it is thought that an anti-Sortlin antibody of the presentdisclosure with the correct epitope specificity can activate the targetantigen, with minimal adverse effects, when the antibody has an Fcdomain from a human IgG2 isotype (CH1 and hinge region) or another typeof Fc domain that is capable of preferentially binding the inhibitoryFcgRIIB r receptors, or a variation thereof.

In some embodiments of any of the antibodies provided herein, themodified antibody Fc is an IgG2 modified Fc. In some embodiments, theIgG2 modified Fc comprises one or more modifications. For example, insome embodiments, the IgG2 modified Fc comprises one or more amino acidsubstitutions (e.g., relative to a wild-type Fc region of the sameisotype). In some embodiments of any of the IgG2 modified Fc, the one ormore amino acid substitutions are selected from V234A (Alegre et al.Transplantation 57:1537-1543 (1994); Xu et al. Cell Immunol, 200:16-26(2000)); G237A (Cole et al. Transplantation, 68:563-571 (1999)); H268Q,V309L, A330S, P331S (US 2007/0148167; Armour et al. Eur J Immunol 29:2613-2624 (1999); Armour et al. The Haematology Journal 1 (Suppl. 1):27(2000); Armour et al. The Haematology Journal 1 (Suppl. 1):27 (2000)),C219S, and/or C220S (White et al. Cancer Cell 27, 138-148 (2015));S267E, L328F (Chu et al. Mol Immunol, 45:3926-3933 (2008)); and M252Y,S254T, and/or T256E according to the EU numbering convention. In someembodiments of any of the IgG2 modified Fc, the Fc comprises an aminoacid substitution at positions V234A and G237A according to EUnumbering. In some embodiments of any of the IgG2 modified Fc, the Fccomprises an amino acid substitution at positions C219S or C220Saccording to EU numbering. In some embodiments of any of the IgG2modified Fc, the Fc comprises an amino acid substitution at positionsA330S and P331S according to EU numbering. In some embodiments of any ofthe IgG2 modified Fc, the Fc comprises an amino acid substitution atpositions S267E and L328F according to EU numbering.

In some embodiments of any of the IgG2 modified Fc, the Fc comprises aC127S amino acid substitution according to the EU numbering convention(White et al., (2015) Cancer Cell 27, 138-148; Lightle et al. ProteinSci. 19:753-762 (2010); and WO 2008/079246). In some embodiments of anyof the IgG2 modified Fc, the antibody has an IgG2 isotype with a Kappalight chain constant domain that comprises a C214S amino acidsubstitution according to the EU numbering convention (White et al.Cancer Cell 27:138-148 (2015); Lightle et al. Protein Sci. 19:753-762(2010); and WO 2008/079246).

In some embodiments of any of the IgG2 modified Fc, the Fc comprises aC220S amino acid substitution according to the EU numbering convention.In some embodiments of any of the IgG2 modified Fc, the antibody has anIgG2 isotype with a Kappa light chain constant domain that comprises aC214S amino acid substitution according to the EU numbering convention.

In some embodiments of any of the IgG2 modified Fc, the Fc comprises aC219S amino acid substitution according to the EU numbering convention.In some embodiments of any of the IgG2 modified Fc, the antibody has anIgG2 isotype with a Kappa light chain constant domain that comprises aC214S amino acid substitution according to the EU numbering convention.

In some embodiments of any of the IgG2 modified Fc, the Fc includes anIgG2 isotype heavy chain constant domain 1 (CH1) and hinge region (Whiteet al. Cancer Cell 27:138-148 (2015)). In certain embodiments of any ofthe IgG2 modified Fc, the IgG2 isotype CH1 and hinge region comprise theamino acid sequence of 118-230 according to EU numbering. In someembodiments of any of the IgG2 modified Fc, the antibody Fc regioncomprises a S267E amino acid substitution, a L328F amino acidsubstitution, or both, and/or a N297A or N297Q amino acid substitutionaccording to the EU numbering convention.

In some embodiments of any of the IgG2 modified Fc, the Fc furthercomprises one or more amino acid substitution at positions E430G, E430S,E430F, E430T, E345K, E345Q, E345R, E345Y, S440Y, and S440W according toEU numbering. In some embodiments of any of the IgG2 modified Fc, the Fcmay further comprise one or more mutations to enhance the antibodyhalf-life in human serum (e.g., one or more (including all) of M252Y,S254T, and T256E mutations according to the EU numbering convention). Insome embodiments of any of the IgG2 modified Fc, the Fc may furthercomprise A330S and P331S.

In some embodiments of any of the IgG2 modified Fc, the Fc is an IgG2/4hybrid Fc. In some embodiments, the IgG2/4 hybrid Fc comprises IgG2 aa118 to 260 and IgG4 aa 261 to 447. In some embodiments of any IgG2modified Fc, the Fc comprises one or more amino acid substitutions atpositions H268Q, V309L, A330S, and P331S according to EU numbering.

In some embodiments of any of the IgG1 and/or IgG2 modified Fc, the Fccomprises one or more additional amino acid substitutions selected fromA330L, L234F; L235E, or P331S according to EU numbering; and anycombination thereof.

In certain embodiments of any of the IgG1 and/or IgG2 modified Fc, theFc comprises one or more amino acid substitutions at a residue positionselected from C127S, L234A, L234F, L235A, L235E, S267E, K322A, L328F,A330S, P331S, E345R, E430G, S440Y, and any combination thereof accordingto EU numbering. In some embodiments of any of the IgG1 and/or IgG2modified Fc, the Fc comprises an amino acid substitution at positionsE430G, L243A, L235A, and P331S according to EU numbering. In someembodiments of any of the IgG1 and/or IgG2 modified Fc, the Fc comprisesan amino acid substitution at positions E430G and P331S according to EUnumbering. In some embodiments of any of the IgG1 and/or IgG2 modifiedFc, the Fc comprises an amino acid substitution at positions E430G andK322A according to EU numbering. In some embodiments of any of the IgG1and/or IgG2 modified Fc, the Fc comprises an amino acid substitution atpositions E430G, A330S, and P331S according to EU numbering. In someembodiments of any of the IgG1 and/or IgG2 modified Fc, the Fc comprisesan amino acid substitution at positions E430G, K322A, A330S, and P331Saccording to EU numbering. In some embodiments of any of the IgG1 and/orIgG2 modified Fc, the Fc comprises an amino acid substitution atpositions E430G, K322A, and A330S according to EU numbering. In someembodiments of any of the IgG1 and/or IgG2 modified Fc, the Fc comprisesan amino acid substitution at positions E430G, K322A, and P331Saccording to EU numbering. In some embodiments of any of the IgG1 and/orIgG2 modified Fc, the Fc comprises an amino acid substitution atpositions S267E and L328F according to EU numbering. In some embodimentsof any of the IgG1 and/or IgG2 modified Fc, the Fc comprises an aminoacid substitution at position C127S according to EU numbering. In someembodiments of any of the IgG1 and/or IgG2 modified Fc, the Fc comprisesan amino acid substitution at positions E345R, E430G and S440Y accordingto EU numbering.

In some embodiments of any of the antibodies provided herein, themodified antibody Fc is an IgG4 modified Fc. In some embodiments, theIgG4 modified Fc comprises one or more modifications. For example, insome embodiments, the IgG4 modified Fc comprises one or more amino acidsubstitutions (e.g., relative to a wild-type Fc region of the sameisotype). In some embodiments of any of the IgG4 modified Fc, the one ormore amino acid substitutions are selected from L235A, G237A, S229P,L236E (Reddy et al. J Immunol 164:1925-1933 (2000)), S267E, E318A,L328F, M252Y, S254T, and/or T256E according to the EU numberingconvention. In some embodiments of any of the IgG4 modified Fc, the Fcmay further comprise L235A, G237A, and E318A according to the EUnumbering convention. In some embodiments of any of the IgG4 modifiedFc, the Fc may further comprise S228P and L235E according to the EUnumbering convention. In some embodiments of any of the IgG4 modifiedFc, the IgG4 modified Fc may further comprise S267E and L328F accordingto the EU numbering convention.

In some embodiments of any of the IgG4 modified Fc, the IgG4 modified Fccomprises may be combined with an S228P mutation according to the EUnumbering convention (Angal et al. Mol Immunol. 30:105-108 (1993))and/or with one or more mutations described in (Peters et al. J BiolChem. 287(29):24525-33 (2012)) to enhance antibody stabilization.

In some embodiments of any of the IgG4 modified Fc, the IgG4 modified Fcmay further comprise one or more mutations to enhance the antibodyhalf-life in human serum (e.g., one or more (including all) of M252Y,S254T, and T256E mutations according to the EU numbering convention).

In some embodiments of any of the IgG4 modified Fc, the Fc comprisesL235E according to EU numbering. In certain embodiments of any of theIgG4 modified Fc, the Fc comprises one or more amino acid substitutionsat a residue position selected from C127S, F234A, L235A, L235E, S267E,K322A, L328F, E345R, E430G, S440Y, and any combination thereof,according to EU numbering. In some embodiments of any of the IgG4modified Fc, the Fc comprises an amino acid substitution at positionsE430G, L243A, L235A, and P331S according to EU numbering. In someembodiments of any of the IgG4 modified Fc, the Fc comprises an aminoacid substitution at positions E430G and P331S according to EUnumbering. In some embodiments of any of the IgG4 modified Fc, the Fccomprises an amino acid substitution at positions E430G and K322Aaccording to EU numbering. In some embodiments of any of the IgG4modified Fc, the Fc comprises an amino acid substitution at positionE430 according to EU numbering. In some embodiments of any of the IgG4modified Fc, the Fc region comprises an amino acid substitution atpositions E430G and K322A according to EU numbering. In some embodimentsof any of the IgG4 modified Fc, the Fc comprises an amino acidsubstitution at positions S267E and L328F according to EU numbering. Insome embodiments of any of the IgG4 modified Fc, the Fc comprises anamino acid substitution at position C127S according to EU numbering. Insome embodiments of any of the IgG4 modified Fc, the Fc comprises anamino acid substitution at positions E345R, E430G and S440Y according toEU numbering.

(9) Other Antibody Modifications

In some embodiments of any of the antibodies, the antibody is aderivative. The term “derivative” refers to a molecule that includes achemical modification other than an insertion, deletion, or substitutionof amino acids (or nucleic acids). In certain embodiments, derivativescomprise covalent modifications, including, but not limited to, chemicalbonding with polymers, lipids, or other organic or inorganic moieties.In certain embodiments, a chemically modified antigen binding proteincan have a greater circulating half-life than an antigen binding proteinthat is not chemically modified. In certain embodiments, a chemicallymodified antigen binding protein can have improved targeting capacityfor desired cells, tissues, and/or organs. In some embodiments, aderivative antigen binding protein is covalently modified to include oneor more water soluble polymer attachments, including, but not limitedto, polyethylene glycol, polyoxyethylene glycol, or polypropyleneglycol. See, e.g., U.S. Pat. Nos. 4,640,835, 4,496,689, 4,301,144,4,670,417, 4,791,192 and 4,179,337. In certain embodiments, a derivativeantigen binding protein comprises one or more polymer, including, butnot limited to, monomethoxy-polyethylene glycol, dextran, cellulose,copolymers of ethylene glycol/propylene glycol, carboxymethylcellulose,polyvinyl pyrrolidone, poly-1,3-dioxolane, poly-1,3,6-trioxane,ethylene/maleic anhydride copolymer, polyaminoacids (either homopolymersor random copolymers), poly-(N-vinyl pyrrolidone)-polyethylene glycol,propylene glycol homopolymers, a polypropylene oxide/ethylene oxideco-polymer, polyoxyethylated polyols (e.g., glycerol) and polyvinylalcohol, as well as mixtures of such polymers.

In certain embodiments, a derivative is covalently modified withpolyethylene glycol (PEG) subunits. In certain embodiments, one or morewater-soluble polymer is bonded at one or more specific position, forexample at the amino terminus, of a derivative. In certain embodiments,one or more water-soluble polymer is randomly attached to one or moreside chains of a derivative. In certain embodiments, PEG is used toimprove the therapeutic capacity for an antigen binding protein. Incertain embodiments, PEG is used to improve the therapeutic capacity fora humanized antibody. Certain such methods are discussed, for example,in U.S. Pat. No. 6,133,426, which is hereby incorporated by referencefor any purpose.

Peptide analogs are commonly used in the pharmaceutical industry asnon-peptide drugs with properties analogous to those of the templatepeptide. These types of non-peptide compound are termed “peptidemimetics” or “peptidomimetics.” Fauchere, J. Adv. Drug Res., 15:29(1986); and Evans et al. J. Med. Chem., 30:1229 (1987), which areincorporated herein by reference for any purpose. Such compounds areoften developed with the aid of computerized molecular modeling. Peptidemimetics that are structurally similar to therapeutically usefulpeptides can be used to produce a similar therapeutic or prophylacticeffect. Generally, peptidomimetics are structurally similar to aparadigm polypeptide (i.e., a polypeptide that has a biochemicalproperty or pharmacological activity), such as human antibody, but haveone or more peptide linkages optionally replaced by a linkage selectedfrom: —CH₂NH—, —CH₂S—,—CH₂—CH₂—,-CH═CH-(cis and trans), —COCH₂—,—CH(OH)CH₂—, and —CH₂SO—, by methods well known in the art. Systematicsubstitution of one or more amino acids of a consensus sequence with aD-amino acid of the same type (e.g., D-lysine in place of L-lysine) canbe used in certain embodiments to generate more stable peptides. Inaddition, constrained peptides comprising a consensus sequence or asubstantially identical consensus sequence variation can be generated bymethods known in the art (Rizo and Gierasch Ann. Rev. Biochem., 61:387(1992), incorporated herein by reference for any purpose); for example,by adding internal cysteine residues capable of forming intramoleculardisulfide bridges which cyclize the peptide.

Drug conjugation involves coupling of a biological active cytotoxic(anticancer) payload or drug to an antibody that specifically targets acertain tumor marker (e.g. a polypeptide that, ideally, is only to befound in or on tumor cells). Antibodies track these proteins down in thebody and attach themselves to the surface of cancer cells. Thebiochemical reaction between the antibody and the target protein(antigen) triggers a signal in the tumor cell, which then absorbs orinternalizes the antibody together with the cytotoxin. After the ADC isinternalized, the cytotoxic drug is released and kills the cancer. Dueto this targeting, ideally the drug has lower side effects and gives awider therapeutic window than other chemotherapeutic agents. Technics toconjugate antibodies are disclosed are known in the art (see, e.g., Janede Lartigue OncLive Jul. 5, 2012; ADC Review on antibody-drugconjugates; and Ducry et al. Bioconjugate Chemistry 21 (1):5-13 (2010).

Binding Assays and Other Assays

Anti-Sortilin antibodies of the present disclosure may be tested forantigen binding activity, e.g., by known methods such as ELISA, surfaceplasmon resonance (SPR), Western blot, flow cytometry, FACS, Bio-layerinterferometry etc.

In some embodiments, competition assays may be used to identify anantibody that competes with any of the antibodies described herein. Insome embodiments, competition assays may be used to identify an antibodythat competes with any of the antibodies listed in Tables 1-30, orcomprising the heavy chain variable region and the light chain variableregion of an antibody selected from S-60, S-60-1, S-60-2, S-60-3,S-60-4, S-60-7, S-60-8, S-60-10, S-60-11, S-60-12, S-60-13, S-60-14,S-60-15 [N33 (wt)], S-60-15.1 [N33T], S-60-15.2 [N33S], S-60-15.3[N33G], S-60-15.4 [N33R], S-60-15.5 [N33D], S-60-15.6 [N33H], S-60-15.7[N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y], S-60-15.10 [N33E],S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14[N33V], S-60-15.15 [N33A], S-60-15.16 [N33M], S-60-15.17 [N33L],S-60-16, S-60-18, S-60-19, and S-60-24 for binding to Sortilin. Incertain embodiments, such a competing antibody binds to the same epitope(e.g., a linear or a conformational epitope) that is bound by any of theantibodies listed in Tables 1-30, or comprising the heavy chain variableregion and the light chain variable region of an antibody selected fromS-60, S-60-1, S-60-2, S-60-3, S-60-4, S-60-7, S-60-8, S-60-10, S-60-11,S-60-12, S-60-13, S-60-14, S-60-15 [N33 (wt)], S-60-15.1 [N33T],S-60-15.2 [N33S], S-60-15.3 [N33G], S-60-15.4 [N33R], S-60-15.5 [N33D],S-60-15.6 [N33H], S-60-15.7 [N33K], S-60-15.8 [N33Q], S-60-15.9 [N33Y],S-60-15.10 [N33E], S-60-15.11 [N33W], S-60-15.12 [N33F], S-60-15.13[N33I], S-60-15.14 [N33V], S-60-15.15 [N33A], S-60-15.16 [N33M],S-60-15.17 [N33L], S-60-16, S-60-18, S-60-19, and S-60-24. Detailedexemplary methods for mapping an epitope to which an antibody binds areprovided in Morris (1996) “Epitope Mapping Protocols,” in Methods inMolecular Biology vol. 66 (Humana Press, Totowa, N.J.).

In an exemplary competition assay, immobilized Sortilin or cellsexpressing Sortilin on a cell surface are incubated in a solutioncomprising a first labeled antibody that binds to Sortilin (e.g., humanor non-human primate) and a second unlabeled antibody that is beingtested for its ability to compete with the first antibody for binding toSortilin. The second antibody may be present in a hybridoma supernatant.As a control, immobilized Sortilin or cells expressing Sortilin isincubated in a solution comprising the first labeled antibody but notthe second unlabeled antibody. After incubation under conditionspermissive for binding of the first antibody to Sortilin, excess unboundantibody is removed, and the amount of label associated with immobilizedSortilin or cells expressing Sortilin is measured. If the amount oflabel associated with immobilized Sortilin or cells expressing Sortilinis substantially reduced in the test sample relative to the controlsample, then that indicates that the second antibody is competing withthe first antibody for binding to Sortilin. See, Harlow and Lane (1988)Antibodies: A Laboratory Manual ch. 14 (Cold Spring Harbor Laboratory,Cold Spring Harbor, N.Y.).

Ligand Binding Assays

Further provided herein are methods of screening for anti-Sortilinantibodies that bind His131, Val132, Pro133, Leu134, Val135, Ile136,Met137, Thr138, Arg196, Phe198, Arg199, Phe203, Lys205, Phe207, Thr210,Thr218, Tyr222, Ser223, Ser227, Ser242, Lys243, Lys248, Lys254, Lys260,Ser305, Phe306, Gly307, Arg311, Phe314, Ser316, Arg325, Arg326, Ile327,Phe350, Tyr351, Ser352, Ile353, Asn373, Ser379, Arg382, Tyr386, Ser595,and/or Glu700 of human Sortilin (SEQ ID NO: 81); or to amino acidresidues of a mammalian Sortilin that corresponds to one or more aminoacid residues His131, Val132, Pro133, Leu134, Val135, Ile136, Met137,Thr138, Arg196, Phe198, Arg199, Phe203, Lys205, Phe207, Thr210, Thr218,Tyr222, Ser223, Ser227, Ser242, Lys243, Lys248, Lys254, Lys260, Ser305,Phe306, Gly307, Arg311, Phe314, Ser316, Arg325, Arg326, Ile327, Phe350,Tyr351, Ser352, Ile353, Asn373, Ser379, Arg382, Tyr386, Ser595, and/orGlu700 of SEQ ID NO: 81, thereby blocking the interactions betweenSortilin and a Sortilin ligand (e.g., Progranulin, pro-neurotrophin,pro-NGF, pro-BDNF, pro-NT3, p75, APP, LpL, APOA5, APOE). In someembodiments, a peptide library can be synthesized in which a Sortilinprotein is dissected into consecutive 15-mer and 25-mer peptidesseparated by one amino acid residue and subsequently spotted ontofilters. Binding of a Sortilin ligand can then tested for its ability tointeract with the receptor peptide or with peptides that are, forexample, mutated at His131, Val132, Pro133, Leu134, Val135, Ile136,Met137, Thr138, Arg196, Phe198, Arg199, Phe203, Lys205, Phe207, Thr210,Thr218, Tyr222, Ser223, Ser227, Ser242, Lys243, Lys248, Lys254, Lys260,Ser305, Phe306, Gly307, Arg311, Phe314, Ser316, Arg325, Arg326, Ile327,Phe350, Tyr351, Ser352, Ile353, Asn373, Ser379, Arg382, Tyr386, Ser595,and/or Glu700 of human Sortilin libraries in the presence or absence ofthe anti-Sortilin antibodies by SPOT binding analysis (e.g., Frank, Rand Overwin, H (1996) Methods. Mol. Biol. 66, 149-169; Reineke, U etal., (2002) J. Immunol. Methods 267, 13-26; and Andersen, O S et al.,(2010) J. Biological Chemistry 285, 12210-12222).

Further provided herein are methods of screening for anti-Sortilinantibodies that block interactions (e.g., binding) Sortilin and aSortilin ligand (e.g., Progranulin, pro-neurotrophin, pro-NGF, pro-BDNF,pro-NT3, p75, APP, LpL, APOA5, APOE). In some embodiments, theinteraction between Sortilin and Sortilin ligands (e.g., Progranulin,pro-neurotrophin, pro-NGF, pro-BDNF, pro-NT3, p75, APP, LpL, APOA5,APOE) may be characterized using surface plasmon resonance analysis(e.g., Skeldal, S et al., (2012) J Biol Chem., 287:43798; and Andersen,O S et al., (2010) The Journal Of Biological Chemistry, 285,12210-12222, a pulldown assay (e.g., Andersen, O S et al., (2010) TheJournal Of Biological Chemistry, 285, 12210-12222, cellulose-boundproteins (e.g., Andersen, O S et al., (2010) The Journal Of BiologicalChemistry, 285, 12210-12222), a proximity ligation assay (e.g.,Gustafsen, C et al., (2013) The Journal of Neuroscience, 33:64-71),and/or alkaline phosphatase-tagged ligands in cell binding assays (e.g.,Hu, F et al., (2010) Neuron 68, 654-667).

Cell-Based Assays

Further provided herein are methods of screening for a Sortilin bindingantagonist, such as an anti-Sortilin antibody, that include contactingan agent (e.g., an anti-Sortilin antibody) with a cell expressing aSortilin protein on its cell surface. In some embodiments, the agent andcell are further contacted with a Sortilin ligand of the presentdisclosure. In some embodiments, the cell itself expresses a Sortilinligand of the present disclosure. The cell-based methods areparticularly suited for screening and validating Sortilin bindingantagonists (e.g., anti-Sortilin antibodies) by assessing the effect onthe interaction between Sortilin and a Sortilin ligand in the context ofa cell.

Accordingly, certain aspects of the present disclosure relate to a cellexpressing a Sortilin protein of the present disclosure on its cellsurface. In some embodiments, the cell endogenously expresses a Sortilinprotein of the present disclosure. In some embodiments, the cell isrecombinantly engineered to express a Sortilin protein of the presentdisclosure. In any of these embodiments, the Sortilin protein of thepresent disclosure (whether endogenous or recombinant) encoded by thepolynucleotide will preferably include at least protein domains requiredfor post-translational processing, membrane translocation, and targetingto the cell surface, including without limitation a signal peptide and atransmembrane domain. In some embodiments, the signal peptide and/ortransmembrane domain may refer to the endogenous Sortilin signal peptideand/or transmembrane domain. In other embodiments, the signal peptideand/or transmembrane domain may refer to an exogenous signal peptideand/or transmembrane domain known to promote cell surface expression inthe desired host cell. In preferred embodiments, the Sortilin proteinwill also contain a domain sufficient for binding a Sortilin ligand ofthe present disclosure.

In these embodiments, any cell that expresses a Sortilin protein of thepresent disclosure on its cell surface may be used. In some embodiments,the cell endogenously expresses a Sortilin protein of the presentdisclosure on its cell surface. In some embodiments, the cell isrecombinantly engineered to express a Sortilin protein of the presentdisclosure on its cell surface. Any suitable Sortilin ligand of thepresent disclosure may be used, such that it retains the ability to bindto the Sortilin protein expressed on the cell surface. The Sortilinligand need not be fluorescently labeled. Levels of Sortilin ligand maybe detected by any assay known in the art, including without limitationELISA, SPR, Western blotting, mass spectrometry, immunoprecipitation,peptide microarray, and so forth.

In some embodiments, the methods disclosed herein involve culturing acell that expresses both a Sortilin protein on its cell surface and aSortilin ligand in a media under conditions in which the Sortilinprotein and the Sortilin ligand are expressed and the Sortilin ligand isreleased into the media; contacting the cell with an agent (e.g., ananti-Sortilin antibody) under conditions in which the Sortilin proteinis capable of binding to the Sortilin ligand; and detecting an increasein the level of the Sortilin ligand in the media, as compared to thelevel of the Sortilin ligand in the media in the absence of the agent.An increase in the level of the Sortilin ligand indicates that the agentis a Sortilin binding antagonist. Without wishing to be bound to theory,it is thought that the interaction between the Sortilin proteinexpressed on the cell surface and the secreted Sortilin ligand willresult in endocytosis and lysosomal degradation of the Sortilin ligand.Therefore, it is thought that decreasing this interaction (e.g., byaddition of a Sortilin binding antagonist of the present disclosure)leads to an increase in the level of the Sortilin ligand in the mediaover time.

In these embodiments, any cell that expresses a Sortilin protein of thepresent disclosure on its cell surface and expresses and secretes aSortilin ligand of the present disclosure may be used. In someembodiments, the cell may endogenously express a Sortilin protein of thepresent disclosure on its cell surface. In some embodiments, the cellmay endogenously express and secrete a Sortilin ligand of the presentdisclosure. In some embodiments, the cell is a U-251 cell, and theSortilin ligand is a Progranulin protein. In some embodiments, the cellmay be recombinantly engineered to express a Sortilin protein of thepresent disclosure on its cell surface. In some embodiments, the cellmay be recombinantly engineered to express and secrete a Sortilin ligandof the present disclosure.

In any of the cell-based assays described herein, a Sortilin ligand ofthe present disclosure may be used. In some embodiments, the Sortilinligand is a Progranulin protein. The Sortilin ligand may be afull-length protein, or it may be a Sortilin-binding peptide fragmentthereof.

Nucleic Acids, Vectors, and Host Cells

Anti-Sortilin antibodies of the present disclosure may be produced usingrecombinant methods and compositions, e.g., as described in U.S. Pat.No. 4,816,567. In some embodiments, isolated nucleic acids having anucleotide sequence encoding any of the anti-Sortilin antibodies of thepresent disclosure are provided. Such nucleic acids may encode an aminoacid sequence comprising the VL and/or an amino acid sequence comprisingthe VH of the anti-Sortilin antibody (e.g., the light and/or heavychains of the antibody). In some embodiments, one or more vectors (e.g.,expression vectors) comprising such nucleic acids are provided. In someembodiments, a host cell comprising such nucleic acid is also provided.In some embodiments, the host cell comprises (e.g., has been transducedwith): (1) a vector comprising a nucleic acid that encodes an amino acidsequence comprising the VL of the antibody and an amino acid sequencecomprising the VH of the antibody, or (2) a first vector comprising anucleic acid that encodes an amino acid sequence comprising the VL ofthe antibody and a second vector comprising a nucleic acid that encodesan amino acid sequence comprising the VH of the antibody. In someembodiments, the host cell is eukaryotic, e.g., a Chinese Hamster Ovary(CHO) cell or lymphoid cell (e.g., Y0, NS0, Sp20 cell). Host cells ofthe present disclosure also include, without limitation, isolated cells,in vitro cultured cells, and ex vivo cultured cells.

Methods of making an anti-Sortilin antibody of the present disclosureare provided. In some embodiments, the method includes culturing a hostcell of the present disclosure comprising a nucleic acid encoding theanti-Sortilin antibody, under conditions suitable for expression of theantibody. In some embodiments, the antibody is subsequently recoveredfrom the host cell (or host cell culture medium).

For recombinant production of an anti-Sortilin antibody of the presentdisclosure, a nucleic acid encoding the anti-Sortilin antibody isisolated and inserted into one or more vectors for further cloningand/or expression in a host cell. Such nucleic acid may be readilyisolated and sequenced using conventional procedures (e.g., by usingoligonucleotide probes that are capable of binding specifically to genesencoding the heavy and light chains of the antibody).

Suitable vectors comprising a nucleic acid sequence encoding any of theanti-Sortilin antibodies of the present disclosure, or cell-surfaceexpressed fragments or polypeptides thereof polypeptides (includingantibodies) described herein include, without limitation, cloningvectors and expression vectors. Suitable cloning vectors can beconstructed according to standard techniques, or may be selected from alarge number of cloning vectors available in the art. While the cloningvector selected may vary according to the host cell intended to be used,useful cloning vectors generally have the ability to self-replicate, maypossess a single target for a particular restriction endonuclease,and/or may carry genes for a marker that can be used in selecting clonescomprising the vector. Suitable examples include plasmids and bacterialviruses, e.g., pUC18, pUC19, Bluescript (e.g., pBS SK+) and itsderivatives, mp18, mp19, pBR322, pMB9, ColE1, pCR1, RP4, phage DNAs, andshuttle vectors such as pSA3 and pAT28. These and many other cloningvectors are available from commercial vendors such as BioRad,Strategene, and Invitrogen.

Suitable host cells for cloning or expression of antibody-encodingvectors include prokaryotic or eukaryotic cells. For example,anti-Sortilin antibodies of the present disclosure may be produced inbacteria, in particular when glycosylation and Fc effector function arenot needed. For expression of antibody fragments and polypeptides inbacteria (e.g., U.S. Pat. Nos. 5,648,237, 5,789,199, and 5,840,523.After expression, the antibody may be isolated from the bacterial cellpaste in a soluble fraction and can be further purified.

In addition to prokaryotes, eukaryotic microorganisms, such asfilamentous fungi or yeast, are also suitable cloning or expressionhosts for antibody-encoding vectors, including fungi and yeast strainswhose glycosylation pathways have been “humanized,” resulting in theproduction of an antibody with a partially or fully human glycosylationpattern (e.g., Gerngross Nat. Biotech. 22:1409-1414 (2004); and Li etal. Nat. Biotech. 24:210-215 (2006)).

Suitable host cells for the expression of glycosylated antibody can alsobe derived from multicellular organisms (invertebrates and vertebrates).Examples of invertebrate cells include plant and insect cells. Numerousbaculoviral strains have been identified which may be used inconjunction with insect cells, particularly for transfection ofSpodoptera frugiperda cells. Plant cell cultures can also be utilized ashosts (e.g., U.S. Pat. Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978,and 6,417,429, describing PLANTIBODIES™ technology for producingantibodies in transgenic plants).

Vertebrate cells may also be used as hosts. For example, mammalian celllines that are adapted to grow in suspension may be useful. Otherexamples of useful mammalian host cell lines are monkey kidney CV1 linetransformed by SV40 (COS-7); human embryonic kidney line (293 or 293cells as described, e.g., in Graham et al. J. Gen Virol. 36:59 (1977));baby hamster kidney cells (BHK); mouse sertoli cells (TM4 cells asdescribed, e.g., in Mather, Biol. Reprod. 23:243-251 (1980)); monkeykidney cells (CV1); African green monkey kidney cells (VERO-76); humancervical carcinoma cells (HELA); canine kidney cells (MDCK; buffalo ratliver cells (BRL 3A); human lung cells (W138); human liver cells (HepG2); mouse mammary tumor (MMT 060562); TRI cells, as described, e.g., inMather et al. Annals N.Y. Acad. Sci. 383:44-68 (1982); MRC 5 cells; andFS4 cells. Other useful mammalian host cell lines include Chinesehamster ovary (CHO) cells, including DHFR− CHO cells (Urlaub et al.Proc. Natl. Acad. Sci. USA 77:4216 (1980)); and myeloma cell lines suchas Y0, NS0 and Sp2/0. For a review of certain mammalian host cell linessuitable for antibody production, see, e.g., Yazaki and Wu, Methods inMolecular Biology, Vol. 248 (B. K. C. Lo, ed., Humana Press, Totowa,N.J.), pp. 255-268 (2003).

Pharmaceutical Compositions

Provided herein are pharmaceutical compositions and/or pharmaceuticalformulations comprising the anti-Sortilin antibodies of the presentdisclosure and a pharmaceutically acceptable carrier.

In some embodiments, pharmaceutically acceptable carrier preferably arenontoxic to recipients at the dosages and concentrations employed. Theantibodies described herein may be formulated into preparations insolid, semi-solid, liquid or gaseous forms. Examples of suchformulations include, without limitation, tablets, capsules, powders,granules, ointments, solutions, suppositories, injections, inhalants,gels, microspheres, and aerosols. Pharmaceutically acceptable carrierscan include, depending on the formulation desired,pharmaceutically-acceptable, non-toxic carriers of diluents, which arevehicles commonly used to formulate pharmaceutical compositions foranimal or human administration. In certain embodiments, thepharmaceutical composition can comprise formulation materials formodifying, maintaining or preserving, for example, the pH, osmolarity,viscosity, clarity, color, isotonicity, odor, sterility, stability, rateof dissolution or release, adsorption or penetration of the composition.

In certain embodiments, pharmaceutically acceptable carriers include,but are not limited to, amino acids (such as glycine, glutamine,asparagine, arginine or lysine); antimicrobials; antioxidants (such asascorbic acid, sodium sulfite or sodium hydrogen-sulfite); buffers (suchas borate, bicarbonate, Tris-HCl, citrates, phosphates or other organicacids); bulking agents (such as mannitol or glycine); chelating agents(such as ethylenediamine tetraacetic acid (EDTA)); complexing agents(such as caffeine, polyvinylpyrrolidone, beta-cyclodextrin orhydroxypropyl-beta-cyclodextrin); fillers; monosaccharides;disaccharides; and other carbohydrates (such as glucose, mannose ordextrins); proteins (such as serum albumin, gelatin or immunoglobulins);coloring, flavoring and diluting agents; emulsifying agents; hydrophilicpolymers (such as polyvinylpyrrolidone); low molecular weightpolypeptides; salt-forming counterions (such as sodium); preservatives(such as benzalkonium chloride, benzoic acid, salicylic acid,thimerosal, phenethyl alcohol, methylparaben, propylparaben,chlorhexidine, sorbic acid or hydrogen peroxide); solvents (such asglycerin, propylene glycol or polyethylene glycol); sugar alcohols (suchas mannitol or sorbitol); suspending agents; surfactants or wettingagents (such as pluronics, PEG, sorbitan esters, polysorbates such aspolysorbate 20, polysorbate 80, triton, tromethamine, lecithin,cholesterol, tyloxapal); stability enhancing agents (such as sucrose orsorbitol); tonicity enhancing agents (such as alkali metal halides,preferably sodium or potassium chloride, mannitol sorbitol); deliveryvehicles; diluents; excipients and/or pharmaceutical adjuvants. Furtherexamples of formulations that are suitable for various types ofadministration can be found in Remington: The Science and Practice ofPharmacy, Pharmaceutical Press 22nd ed. (2013). For a brief review ofmethods for drug delivery, see, Langer, Science 249:1527-1533 (1990).

Formulations suitable for parenteral administration include aqueous andnon-aqueous, isotonic sterile injection solutions, which can compriseantioxidants, buffers, bacteriostats, and solutes that render theformulation isotonic with the blood of the intended recipient, andaqueous and non-aqueous sterile suspensions that can include suspendingagents, solubilizers, thickening agents, stabilizers, and preservatives.

Formulations may be optimized for retention and stabilization in thebrain or central nervous system. When the agent is administered into thecranial compartment, it is desirable for the agent to be retained in thecompartment, and not to diffuse or otherwise cross the blood brainbarrier. Stabilization techniques include cross-linking, multimerizing,or linking to groups such as polyethylene glycol, polyacrylamide,neutral protein carriers, etc. in order to achieve an increase inmolecular weight.

Other strategies for increasing retention include the entrapment of theantibody, such as an anti-Sortilin antibody of the present disclosure,in a biodegradable or bioerodible implant. The rate of release of thetherapeutically active agent is controlled by the rate of transportthrough the polymeric matrix, and the biodegradation of the implant.Implants may be particles, sheets, patches, plaques, fibers,microcapsules and the like and may be of any size or shape compatiblewith the selected site of insertion. Biodegradable polymericcompositions which may be employed may be organic esters or ethers,which when degraded result in physiologically acceptable degradationproducts, including the monomers. Anhydrides, amides, orthoesters or thelike, by themselves or in combination with other monomers, may find use.The polymers will be condensation polymers. The polymers may becross-linked or non-cross-linked. Of particular interest are polymers ofhydroxyaliphatic carboxylic acids, either homo- or copolymers, andpolysaccharides. Included among the polyesters of interest are polymersof D-lactic acid, L-lactic acid, racemic lactic acid, glycolic acid,polycaprolactone, and combinations thereof. Among the polysaccharides ofinterest are calcium alginate, and functionalized celluloses,particularly carboxymethylcellulose esters characterized by being waterinsoluble, a molecular weight of about 5 kD to 500 kD, etc.Biodegradable hydrogels may also be employed in the implants of thesubject invention. Hydrogels are typically a copolymer material,characterized by the ability to imbibe a liquid.

Pharmaceutical Dosages

An antibody provided herein (and any additional therapeutic agent) canbe administered by any suitable means, including parenteral,intrapulmonary, intranasal, intralesional administration,intracerobrospinal, intracranial, intraspinal, intrasynovial,intrathecal, oral, topical, or inhalation routes. Parenteral infusionsinclude intramuscular, intravenous administration as a bolus or bycontinuous infusion over a period of time, intraarterial,intra-articular, intraperitoneal, or subcutaneous administration. Insome embodiments, the administration is intravenous administration. Insome embodiments, the administration is subcutaneous. Dosing can be byany suitable route, e.g. by injections, such as intravenous orsubcutaneous injections, depending in part on whether the administrationis brief or chronic. Various dosing schedules including but not limitedto single or multiple administrations over various time-points, bolusadministration, and pulse infusion are contemplated herein.

Antibodies provided herein would be formulated, dosed, and administeredin a fashion consistent with good medical practice. Factors forconsideration in this context include the particular disorder beingtreated, the particular mammal being treated, the clinical condition ofthe individual patient, the cause of the disorder, the site of deliveryof the agent, the method of administration, the scheduling ofadministration, and other factors known to medical practitioners. Theantibody need not be, but is optionally formulated with one or moreagents currently used to prevent or treat the disorder in question. Theeffective amount of such other agents depends on the amount of antibodypresent in the formulation, the type of disorder or treatment, and otherfactors discussed above. These are generally used in the same dosagesand with administration routes as described herein, or about from 1 to99% of the dosages described herein, or in any dosage and by any routethat is empirically/clinically determined to be appropriate.

Dosages for a particular anti-Sortilin antibody may be determinedempirically in individuals who have been given one or moreadministrations of the anti-Sortilin antibody. Individuals are givenincremental doses of an anti-Sortilin antibody. To assess efficacy of ananti-Sortilin antibody, a clinical symptom of any of the diseases,disorders, or conditions of the present disclosure (e.g., frontotemporaldementia, Alzheimer's disease, vascular dementia, seizures, retinaldystrophy, a traumatic brain injury, a spinal cord injury, long-termdepression, atherosclerotic vascular diseases, and undesirable symptomsof normal aging) can be monitored.

For the prevention or treatment of disease, the appropriate dosage of anantibody of the invention (when used alone or in combination with one ormore other additional therapeutic agents) will depend on the type ofdisease to be treated, the type of antibody, the severity and course ofthe disease, whether the antibody is administered for preventive ortherapeutic purposes, previous therapy, the patient's clinical historyand response to the antibody, and the discretion of the attendingphysician. The antibody is suitably administered to the patient at onetime or over a series of treatments.

Depending on the type and severity of the disease, about 1 μg/kg to 15mg/kg (e.g., 0.1 mg/kg-10 mg/kg) of antibody can be an initial candidatedosage for administration to the patient, whether, for example, by oneor more separate administrations, or by continuous infusion. One typicaldaily dosage might range from about 1 μg/kg to 100 mg/kg or more,depending on the factors mentioned above. For repeated administrationsover several days or longer, depending on the condition, the treatmentwould generally be sustained until a desired suppression of diseasesymptoms occurs. One exemplary dosage of the antibody would be in therange from about 0.05 mg/kg to about 10 mg/kg. Thus, one or more dosesof about 0.5 mg/kg, 2.0 mg/kg, 4.0 mg/kg or 10 mg/kg (or any combinationthereof) may be administered to the patient. Such doses may beadministered intermittently, e.g., every week or every three weeks(e.g., such that the patient receives from about two to about twenty, ore.g., about six doses of the antibody). In certain embodiments, dosingfrequency is three times per day, twice per day, once per day, onceevery other day, once weekly, once every two weeks, once every fourweeks, once every five weeks, once every six weeks, once every sevenweeks, once every eight weeks, once every nine weeks, once every tenweeks, or once monthly, once every two months, once every three months,or longer. An initial higher loading dose, followed by one or more lowerdoses may be administered. However, other dosage regimens may be useful.The progress of this therapy is easily monitored by conventionaltechniques and assays.

Therapeutic Uses

In certain aspects, anti-Sortilin antibodies of the present disclosurecan be used for preventing, reducing risk for, or treating an individualhaving a disease, disorder, or injury. Anti-Sortilin antibodies of thepresent disclosure can be used to prevent, reduce risk of, or treat celldeath (e.g., neuronal cell death), frontotemporal dementia, Alzheimer'sdisease, vascular dementia, seizures, retinal dystrophy, a traumaticbrain injury, a spinal cord injury, long-term depression,atherosclerotic vascular diseases, undesirable symptoms of normal aging,dementia, mixed dementia, Creutzfeldt-Jakob disease, normal pressurehydrocephalus, amyotrophic lateral sclerosis, Huntington's disease,taupathy disease, stroke, acute trauma, chronic trauma, lupus, acute andchronic colitis, Crohn's disease, inflammatory bowel disease, ulcerativecolitis, malaria, essential tremor, central nervous system lupus,Behcet's disease, Parkinson's disease, dementia with Lewy bodies,multiple system atrophy, intervertebral disc degeneration, Shy-Dragersyndrome, progressive supranuclear palsy, cortical basal ganglionicdegeneration, acute disseminated encephalomyelitis, granulomartousdisorders, Sarcoidosis, diseases of aging, age related maculardegeneration, glaucoma, retinitis pigmentosa, retinal degeneration,respiratory tract infection, sepsis, eye infection, systemic infection,inflammatory disorders, arthritis, multiple sclerosis, metabolicdisorder, obesity, insulin resistance, type 2 diabetes, tissue orvascular damage, an injury, and/or one or more undesirable symptoms ofnormal aging.

In certain aspects, provided herein is a method of preventing, reducingrisk for, or treating an individual having a disease, disorder, orinjury, comprising administering to an individual in need thereof atherapeutically effective amount of an anti-Sortilin antibody of thepresent disclosure. In some embodiments, the disease, disorder or injuryis selected from the group consisting of frontotemporal dementia,progressive supranuclear palsy, Alzheimer's disease, vascular dementia,seizures, retinal dystrophy, amyotrophic lateral sclerosis, traumaticbrain injury, a spinal cord injury, dementia, stroke, Parkinson'sdisease, acute disseminated encephalomyelitis, retinal degeneration, agerelated macular degeneration, glaucoma, multiple sclerosis, septicshock, bacterial infection, arthritis, and osteoarthritis.

Certain aspects of the present disclosure provide methods of increasingProgranulin levels in an individual in need thereof, such as in thebrain, blood, and/or peripheral organs of the individual, byadministering to the individual a therapeutically effective amount ofone or more anti-Sortilin antibodies of the present disclosure. Otheraspects of the present disclosure provide methods of increasingextracellular levels of Progranulin, by contacting one or more cellswith one or more anti-Sortilin antibodies of the present disclosure. Insome embodiments, levels of Progranulin are increased without decreasingcellular levels of Sortilin. Other aspects of the present disclosureprovide methods of decreasing cellular levels of Sortilin in anindividual in need thereof, such as in the brain and/or peripheralorgans of the individual, by administering to the individual atherapeutically effective amount of one or more anti-Sortilin antibodiesof the present disclosure. Other aspects of the present disclosureprovide methods of decreasing cellular levels of Sortilin of one or morecells, comprising contacting one or more cells with one or moreanti-Sortilin antibodies of the present disclosure.

Further aspects of the present disclosure provide methods for increasingthe effective concentrations of Progranulin and/or reducing theeffective concentrations of a neurotrophin of the present disclosure(e.g., pro-neurotrophin-3, pro-neurotrophin-4/5, pro-neurotrophins,pro-NGF, pro-BDNF, neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.),neurotensin, p75, Sortilin propeptide (Sort-pro), amyloid precursorprotein (APP), the A beta peptide, lipoprotein lipase (LpL),apolipoprotein AV (APOA5), apolipoprotein E (APOE), PCSK9, and receptorassociated protein (RAP) in an individual in need thereof, byadministering to the individual a therapeutically effective amount of ananti-Sortilin antibody of the present disclosure to inhibit theinteraction between Sortilin and Progranulin, a neurotrophin of thepresent disclosure (e.g., pro-neurotrophins, pro-neurotrophin-3,pro-neurotrophin-4/5, pro-NGF, pro-BDNF, neurotrophin-3,neurotrophin-4/5, NGF, BDNF, etc.), neurotensin, p75, Sortilinpropeptide (Sort-pro), amyloid precursor protein (APP), the A betapeptide, lipoprotein lipase (LpL), apolipoprotein AV (APOA5),apolipoprotein E (APOE), and/or receptor associated protein (RAP).

The present disclosure also provides methods of inhibiting theinteraction between Sortilin and Progranulin, a neurotrophin of thepresent disclosure (e.g., pro-neurotrophins, pro-neurotrophin-3,pro-neurotrophin-4/5, pro-NGF, pro-BDNF, neurotrophin-3,neurotrophin-4/5, NGF, BDNF, etc.), neurotensin, p75, Sortilinpropeptide (Sort-pro), amyloid precursor protein (APP), the A betapeptide, lipoprotein lipase (LpL), apolipoprotein AV (APOA5),apolipoprotein E (APOE), PCSK9, and/or receptor associated protein(RAP); as well as one or more activities of Sortilin, Progranulin, aneurotrophin of the present disclosure (e.g., pro-neurotrophins,pro-neurotrophin-3, pro-neurotrophin-4/5, pro-NGF, pro-BDNF,neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.), neurotensin, p75,Sortilin propeptide (Sort-pro), amyloid precursor protein (APP), the Abeta peptide, lipoprotein lipase (LpL), apolipoprotein AV (APOA5),apolipoprotein E (APOE), and/or receptor associated protein (RAP) in anindividual by administering to the individual a therapeuticallyeffective amount of an anti-Sortilin antibody of the present disclosure.

In certain aspects, provided herein is a method of preventing, reducingrisk for, or treating an individual having a disease, disorder, orinjury, comprising administering to an individual in need thereof atherapeutically effective amount of an anti-Sortilin antibody of thepresent disclosure.

As disclosed herein, anti-Sortilin antibodies of the present disclosuremay be used for preventing, reducing risk, or treating frontotemporaldementia, Alzheimer's disease, vascular dementia, seizures, retinaldystrophy, a traumatic brain injury, a spinal cord injury, long-termdepression, atherosclerotic vascular diseases, undesirable symptoms ofnormal aging, dementia, mixed dementia, Creutzfeldt-Jakob disease,normal pressure hydrocephalus, amyotrophic lateral sclerosis,Huntington's disease, taupathy disease, stroke, acute trauma, chronictrauma, lupus, acute and chronic colitis, Crohn's disease, inflammatorybowel disease, ulcerative colitis, malaria, essential tremor, centralnervous system lupus, Behcet's disease, Parkinson's disease, dementiawith Lewy bodies, multiple system atrophy, intervertebral discdegeneration, Shy-Drager syndrome, progressive supranuclear palsy,cortical basal ganglionic degeneration, acute disseminatedencephalomyelitis, granulomartous disorders, Sarcoidosis, diseases ofaging, age related macular degeneration, glaucoma, retinitis pigmentosa,retinal degeneration, respiratory tract infection, sepsis, eyeinfection, systemic infection, inflammatory disorders, arthritis,multiple sclerosis, metabolic disorder, obesity, insulin resistance,type 2 diabetes, tissue or vascular damage, an injury, and one or moreundesirable symptoms of normal aging.

In some embodiments, the present disclosure provides methods ofpreventing, reducing risk, or treating an individual havingfrontotemporal dementia, Alzheimer's disease, vascular dementia,seizures, retinal dystrophy, a traumatic brain injury, a spinal cordinjury, long-term depression, atherosclerotic vascular diseases,undesirable symptoms of normal aging, dementia, mixed dementia,Creutzfeldt-Jakob disease, normal pressure hydrocephalus, amyotrophiclateral sclerosis, Huntington's disease, taupathy disease, stroke, acutetrauma, chronic trauma, lupus, acute and chronic colitis, Crohn'sdisease, inflammatory bowel disease, ulcerative colitis, malaria,essential tremor, central nervous system lupus, Behcet's disease,Parkinson's disease, dementia with Lewy bodies, multiple system atrophy,intervertebral disc degeneration, Shy-Drager syndrome, progressivesupranuclear palsy, cortical basal ganglionic degeneration, acutedisseminated encephalomyelitis, granulomartous disorders, Sarcoidosis,diseases of aging, age related macular degeneration, glaucoma, retinitispigmentosa, retinal degeneration, respiratory tract infection, sepsis,eye infection, systemic infection, inflammatory disorders, arthritis,multiple sclerosis, metabolic disorder, obesity, insulin resistance,type 2 diabetes, tissue or vascular damage, an injury, and one or moreundesirable symptoms of normal aging, in an individual in need thereofby administering to the individual a therapeutically effective amount ofan anti-Sortilin antibody of the present disclosure to: (i) inhibit theinteraction between Sortilin and Progranulin, a neurotrophin of thepresent disclosure (e.g., pro-neurotrophins, pro-neurotrophin-3,pro-neurotrophin-4/5, pro-NGF, pro-BDNF, neurotrophin-3,neurotrophin-4/5, NGF, BDNF, etc.), neurotensin, p75, Sortilinpropeptide (Sort-pro), amyloid precursor protein (APP), the A betapeptide, lipoprotein lipase (LpL), apolipoprotein AV (APOA5),apolipoprotein E (APOE), and/or receptor associated protein (RAP);and/or (ii) inhibit one or more activities of Sortilin, Progranulin, aneurotrophin of the present disclosure (e.g., pro-neurotrophins,pro-neurotrophin-3, pro-neurotrophin-4/5, pro-NGF, pro-BDNF,neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.), neurotensin, p75,Sortilin propeptide (Sort-pro), amyloid precursor protein (APP), the Abeta peptide, lipoprotein lipase (LpL), apolipoprotein AV (APOA5),apolipoprotein E (APOE), and/or receptor associated protein (RAP). Insome embodiments, the present disclosure provides methods of inducingwound healingin an individual in need thereof by administering to theindividual a therapeutically effective amount of an anti-Sortilinantibody of the present disclosure.

The present disclosure also provides methods of promoting cell survival,such as neuronal cell survival, by administering an anti-Sortilinantibody of the present disclosure to inhibit the interaction betweenSortilin and Progranulin, a neurotrophin of the present disclosure(e.g., pro-neurotrophins, pro-neurotrophin-3, pro-neurotrophin-4/5,pro-NGF, pro-BDNF, neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.),neurotensin, p75, amyloid precursor protein (APP), and the A betapeptide. The anti-Sortilin antibody may be administered to cells invitro to promote cell survival. Alternatively, the anti-Sortilinantibody may be administered in vivo (e.g., by administering theantibody to an individual) to promote cell survival.

In certain aspects, provided herein is a method of inhibiting one ormore of neuroinflammation, axonopathy characterized by short axonaloutgrowth and aberrant branching, microglial activation, andinflammatory response, comprising administering to the individual atherapeutically effective amount of an anti-Sortilin antibody of thepresent disclosure.

The present disclosure provides methods of inhibiting neuroinflammation,axonopathy characterized by short axonal outgrowth and aberrantbranching, microglial activation, and inflammatory response andpromoting wound healing, autophagy and the clearance of aggregateproteins by administering an anti-Sortilin antibody of the presentdisclosure to inhibit the interaction between Sortilin and Progranulin,a neurotrophin of the present disclosure (e.g., pro-neurotrophins,pro-neurotrophin-3, pro-neurotrophin-4/5, pro-NGF, pro-BDNF,neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.), neurotensin, p75,amyloid precursor protein (APP), and the A beta peptide. Theanti-Sortilin antibody may be administered to cells in vitro.Alternatively, the anti-Sortilin antibody may be administered in vivo(e.g., by administering the antibody to an individual).

In certain aspects, provided herein is a method of promoting one or moreof wound healing, autophagy, and clearance of aggregate proteins,comprising administering to the individual a therapeutically effectiveamount of an anti-Sortilin antibody of the present disclosure.

In certain aspects, provided herein is a method of preventing, reducingrisk, or treating an individual having arthritis, comprisingadministering to the individual a therapeutically effective amount of ananti-Sortilin antibody of the present disclosure.

The present disclosure also provides methods of decreasing expression ofone or more pro-inflammatory mediators by administering to an individualin need thereof an anti-Sortilin antibody of the present disclosure. Insome embodiments, the one or more pro-inflammatory mediators areselected from IL-6, IL12p70, IL12p40, IL-1β, TNF-α, CXCL1, CCL2, CCL3,CCL4, and CCL5.

In some embodiments, a method of the present disclosure includes ananti-Sortilin antibody comprising two or more anti-Sortilin antibodies.

Dementia

Dementia is a non-specific syndrome (i.e., a set of signs and symptoms)that presents as a serious loss of global cognitive ability in apreviously unimpaired person, beyond what might be expected from normalageing. Dementia may be static as the result of a unique global braininjury. Alternatively, dementia may be progressive, resulting inlong-term decline due to damage or disease in the body. While dementiais much more common in the geriatric population, it can also occurbefore the age of 65. Cognitive areas affected by dementia include,without limitation, memory, attention span, language, and problemsolving. Generally, symptoms must be present for at least six months tobefore an individual is diagnosed with dementia.

Exemplary forms of dementia include, without limitation, frontotemporaldementia, Alzheimer's disease, vascular dementia, semantic dementia, anddementia with Lewy bodies.

Without wishing to be bound by theory, it is believed that administeringan anti-Sortilin antibody of the present disclosure can prevent, reducethe risk, and/or treat dementia. In some embodiments, administering ananti-Sortilin antibody may induce one or more Progranulin activities inan individual having dementia (e.g., neurotrophic and/or survivalactivity on neurons, and anti-inflammatory activity.

Frontotemporal Dementia

Frontotemporal dementia (FTD) is a condition resulting from theprogressive deterioration of the frontal lobe of the brain. Over time,the degeneration may advance to the temporal lobe. Second only toAlzheimer's disease (AD) in prevalence, FTD accounts for 20% ofpre-senile dementia cases. The clinical features of FTD include memorydeficits, behavioral abnormalities, personality changes, and languageimpairments (Cruts, M. & Van Broeckhoven, C., Trends Genet. 24:186-194(2008); Neary, D., et al., Neurology 51:1546-1554 (1998); Ratnavalli,E., Brayne, C., Dawson, K. & Hodges, J. R., Neurology 58:1615-1621(2002)).

A substantial portion of FTD cases are inherited in an autosomaldominant fashion, but even in one family, symptoms can span a spectrumfrom FTD with behavioral disturbances, to Primary Progressive Aphasia,to Cortico-Basal Ganglionic Degeneration. FTD, like mostneurodegenerative diseases, can be characterized by the pathologicalpresence of specific protein aggregates in the diseased brain.Historically, the first descriptions of FTD recognized the presence ofintraneuronal accumulations of hyperphosphorylated Tau protein inneurofibrillary tangles or Pick bodies. A causal role for themicrotubule associated protein Tau was supported by the identificationof mutations in the gene encoding the Tau protein in several families(Hutton, M., et al., Nature 393:702-705 (1998). However, the majority ofFTD brains show no accumulation of hyperphosphorylated Tau but doexhibit immunoreactivity to ubiquitin (Ub) and TAR DNA binding protein(TDP43) (Neumann, M., et al., Arch. Neurol. 64:1388-1394 (2007)). Amajority of those FTD cases with Ub inclusions (FTD-U) were shown tocarry mutations in the Progranulin gene.

Progranulin mutations result in haploinsufficiency and are known to bepresent in nearly 50% of familial FTD cases, making Progranulin mutationa major genetic contributor to FTD. Without wishing to be bound bytheory, it is believed that the loss-of-function heterozygous characterof Progranulin mutations indicates that in healthy individuals,Progranulin expression plays a dose-dependent, critical role inprotecting healthy individuals from the development of FTD. Accordingly,increasing levels of Progranulin by inhibiting the interaction betweenSortilin and Progranulin, can prevent, reduce the risk, and/or treatFTD.

In some embodiments, administering an anti-Sortilin antibody of thepresent disclosure, can prevent, reduce the risk, and/or treat FTD. Insome embodiments, administering an anti-Sortilin antibody may modulateone or more Sortilin activities in an individual having FTD.

Alzheimer's Disease

Alzheimer's disease (AD) is the most common form of dementia. There isno cure for the disease, which worsens as it progresses, and eventuallyleads to death. Most often, AD is diagnosed in people over 65 years ofage. However, the less-prevalent early-onset Alzheimer's can occur muchearlier.

Common symptoms of Alzheimer's disease include, behavioral symptoms,such as difficulty in remembering recent events; cognitive symptoms,confusion, irritability and aggression, mood swings, trouble withlanguage, and long-term memory loss. As the disease progresses bodilyfunctions are lost, ultimately leading to death. Alzheimer's diseasedevelops for an unknown and variable amount of time before becomingfully apparent, and it can progress undiagnosed for years.

It has been shown that Sortilin binds to amyloid precursor protein (APP)and the APP processing enzyme BACE1. Without wishing to be bound bytheory, it is believed that these interactions are involved inAlzheimer's disease. Accordingly, and without wishing to be bound bytheory, it is believed that anti-Sortilin antibodies of the presentdisclosure can be utilized to inhibit such interactions and prevent,reduce the risk of, or treat Alzheimer's disease in individuals in needthereof.

In some embodiments, and without wishing to be bound by theory, it isbelieved that anti-Sortilin antibodies of the present disclosure thatinhibit the interaction between Sortilin and neurotrophins of thepresent disclosure (e.g., pro-neurotrophins, pro-neurotrophin-3,pro-neurotrophin-4/5, pro-NGF, pro-BDNF, neurotrophin-3,neurotrophin-4/5, NGF, BDNF, etc.), p75, amyloid precursor protein(APP), and/or the A beta peptide, or that inhibit one or more activitiesof Sortilin can be utilized to prevent, reduce the risk of, or treatAlzheimer's disease in individuals in need thereof.

In some embodiments, administering an anti-Sortilin antibody of thepresent disclosure can prevent, reduce the risk, and/or treatAlzheimer's disease. In some embodiments, administering an anti-Sortilinantibody may modulate one or more Sortilin activities in an individualhaving Alzheimer's disease.

Vascular Dementia

Vascular dementia (VaD) is a subtly progressive worsening of memory andother cognitive functions that is believed to be due to cerebrovasculardisease (vascular disease within the brain). Cerebrovascular disease isthe progressive change in our blood vessels (vasculature) in the brain(cerebrum). The most common vascular change associated with age is theaccumulation of cholesterol and other substances in the blood vesselwalls. This results in the thickening and hardening of the walls, aswell as narrowing of the vessels, which can result in a reduction oreven a complete stopping of blood flow to brain regions supplied by theaffected artery. Vascular dementia patients often present with similarsymptoms to Alzheimer's disease (AD) patients. However, the relatedchanges in the brain are not due to AD pathology but to chronic reducedblood flow in the brain, eventually resulting in dementia. VaD isconsidered one of the most common types of dementia in older adults.Symptoms of VaD include difficulties with memory, difficulty withorganization and solving complex problems, slowed thinking, distractionor “absent mindedness,” difficulty retrieving words from memory, changesin mood or behavior such as depression, irritability, or apathy, andhallucinations or delusions.

Without wishing to be bound by theory, it is believed that one or moreactivities of Sortilin, or one or more interactions between Sortilin andProgranulin, neurotrophins of the present disclosure (e.g.,pro-neurotrophins, pro-neurotrophin-3, pro-neurotrophin-4/5, pro-NGF,pro-BDNF, neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.),neurotensin, lipoprotein lipase, apolipoprotein AV, and/orreceptor-associated protein are involved in vascular dementia.Accordingly, and without wishing to be bound by theory, it is believedthat anti-Sortilin antibodies of the present disclosure that inhibit theinteraction between Sortilin and neurotrophins of the present disclosure(e.g., pro-neurotrophins, pro-neurotrophin-3, pro-neurotrophin-4/5,pro-NGF, pro-BDNF, neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.),neurotensin, p75, Sortilin propeptide (Sort-pro), amyloid precursorprotein (APP), the A beta peptide, lipoprotein lipase (LpL),apolipoprotein AV (APOA5), apolipoprotein E (APOE), and/or receptorassociated protein (RAP); or that inhibit one or more activities ofSortilin can be utilized to prevent, reduce the risk of, or treatvascular dementia in individuals in need thereof.

In some embodiments, administering an anti-Sortilin antibody of thepresent disclosure can prevent, reduce the risk, and/or treat VaD. Insome embodiments, administering an anti-Sortilin antibody may modulateone or more Sortilin activities in an individual having VaD.

Seizures, Retinal Dystrophy, Traumatic Brain Injuries, Spinal CordInjuries, and Long-Term Depression

As used herein, retinal dystrophy refers to any disease or conditionthat involves the degeneration of the retinal. Such diseases orconditions may lead to loss of vision or complete blindness.

As used herein, seizures also include epileptic seizures, and refer to atransient symptom of abnormal excessive or synchronous neuronal activityin the brain. The outward effect can be as dramatic as a wild thrashingmovement or as mild as a brief loss of awareness. Seizures can manifestas an alteration in mental state, tonic or clonic movements,convulsions, and various other psychic symptoms.

Traumatic brain injuries (TBI), may also be known as intracranialinjuries. Traumatic brain injuries occur when an external forcetraumatically injures the brain. Traumatic brain injuries can beclassified based on severity, mechanism (closed or penetrating headinjury), or other features (e.g., occurring in a specific location orover a widespread area).

Spinal cord injuries (SCI) include any injury to the spinal cord that iscaused by trauma instead of disease. Depending on where the spinal cordand nerve roots are damaged, the symptoms can vary widely, from pain toparalysis to incontinence. Spinal cord injuries are described at variouslevels of “incomplete”, which can vary from having no effect on thepatient to a “complete” injury which means a total loss of function.

Long-term depression (LTD) is an activity-dependent reduction in theefficacy of neuronal synapses lasting hours or longer following a longpatterned stimulus. Long-term depression can occur in many areas of thecentral nervous system with varying mechanisms depending upon brainregion and developmental progress. Long-term depression can occur in thehippocampus, cerebellum, and in different types of neurons that releasevarious neurotransmitters. Without wishing to be bound by theory, it isbelieved that long-term depression may be associated withneurodegeneration, dementia, and Alzheimer's disease.

It has been shown that pro-neurotrophins (e.g., pro-neurotrophin-4/5,neurotrophin-4/5, pro-NGF, pro-BDNF, etc.) play a role in seizures,retinal dystrophy, traumatic brain injury, spinal cord injury, andlong-term depression.

Accordingly, and without wishing to be bound by theory, it is believedthat anti-Sortilin antibodies of the present disclosure that inhibit theinteraction between Sortilin and neurotrophins of the present disclosure(e.g., pro-neurotrophins, pro-neurotrophin-3, pro-neurotrophin-4/5,pro-NGF, pro-BDNF, neurotrophin-3, neurotrophin-4/5, NGF, BDNF, etc.);or that inhibit one or more activities of Sortilin can be utilized toprevent, reduce the risk of, or treat seizures, retinal dystrophy,traumatic brain injuries, spinal cord injuries, and/or long-termdepression in individuals in need thereof.

In some embodiments, administering an anti-Sortilin antibody of thepresent disclosure can prevent, reduce the risk, and/or treat seizures,retinal dystrophy, traumatic brain injuries, spinal cord injuries,and/or long-term depression. In some embodiments, administering ananti-Sortilin antibody may modulate one or more Sortilin activities inan individual having seizures, retinal dystrophy, traumatic braininjuries, spinal cord injuries, and/or long-term depression.

Atherosclerotic Vascular Diseases

As used herein, “atherosclerotic vascular disease,” “ASVD,” and“atherosclerosis” are used interchangeably and refer to any condition inwhich an artery wall thickens as a result of the accumulation of fattymaterials such as cholesterol, lipids, and triglyceride. Atheroscleroticvascular diseases include, without limitation, any ASVD-associatedcondition, disorder, or disease, including without limitation,thromboembolism, stroke, ischemia, infarctions, coronary thrombosis,myocardial infarction (e.g., heart attack), and claudication.

As disclosed herein, Sortilin proteins of the present disclosure areinvolved in lipid regulation, by binding lipid-associated proteins, suchas receptor associated protein, lipoprotein lipase and apopolipoproteinsAPOA5 and APOE.

Accordingly, and without wishing to be bound by theory, it is believedthat anti-Sortilin antibodies of the present disclosure that inhibit theinteraction between Sortilin and lipoprotein lipase (LpL),apolipoprotein AV (APOA5), apolipoprotein E (APOE), and/or receptorassociated protein (RAP); or that inhibit one or more activities ofSortilin can be utilized to prevent, reduce the risk of, or treat one ormore atherosclerotic vascular disease in individuals in need thereof.

In some embodiments, administering an anti-Sortilin antibody of thepresent disclosure can prevent, reduce the risk, and/or treatatherosclerotic vascular disease. In some embodiments, administering ananti-Sortilin antibody may modulate one or more Sortilin activities inan individual having atherosclerotic vascular disease.

Undesirable Symptoms of Aging

As used herein, undesirable symptoms of aging include, withoutlimitation, memory loss, behavioral changes, dementia, Alzheimer'sdisease, retinal degeneration, atherosclerotic vascular diseases,hearing loss, and cellular break-down.

In some embodiments, and without wishing to be bound by theory, it isbelieved that anti-Sortilin antibodies of the present disclosure thatinhibit the interaction between Sortilin and Progranulin, neurotrophinsof the present disclosure (e.g., pro-neurotrophins, pro-neurotrophin-3,pro-neurotrophin-4/5, pro-NGF, pro-BDNF, neurotrophin-3,neurotrophin-4/5, NGF, BDNF, etc.), neurotensin, p75, lipoprotein lipase(LpL), apolipoprotein AV (APOA5), and/or receptor associated protein(RAP); or that inhibit one or more activities of Sortilin can beutilized to prevent, reduce the risk of, or treat one or moreundesirable symptoms of aging.

In some embodiments, administering an anti-Sortilin antibody of thepresent disclosure can prevent, reduce the risk, and/or treat one ormore undesirable symptoms of aging. In some embodiments, administeringan anti-Sortilin antibody may modulate one or more Sortilin activitiesin an individual having one or more undesirable symptoms of aging.

Amyotrophic Lateral Sclerosis (ALS)

As used herein, amyotrophic lateral sclerosis (ALS) or, motor neurondisease or, Lou Gehrig's disease are used interchangeably and refer to adebilitating disease with varied etiology characterized by rapidlyprogressive weakness, muscle atrophy and fasciculations, musclespasticity, difficulty speaking (dysarthria), difficulty swallowing(dysphagia), and difficulty breathing (dyspnea).

PGRN haploinsufficiency due to heterozygous loss-of-function mutationsin the GRN gene results in a reduction of CSF PGRN levels and is causalfor the development of frontotemporal dementia (FTD) with TDP-43pathology (Sleegers et al., (2009) Ann Neurol 65:603; Smith et al.,(2012) Am J Hum Genet 90:1102). TDP-43 has also been identified as amajor pathological protein in ALS, suggesting a similarity between ALSand FTD.

For example, over twenty dominant mutations in TDP-43 have beenidentified in sporadic and familial ALS patients (Lagier-Tourenne etal., (2009) Cell 136:1001) and TDP-43 positive aggregates are found inapproximately 95% of ALS cases (Prasad et al., (2019) Front Mol Neurosci12:25). Furthermore, ALS risk genes, such as MOBP, C9ORF72, MOBKL2B, NSFand FUS, can also cause FTD (Karch et al., (2018) JAMA Neurol 75:860).In addition, both PGRN and C9ORF72 mutations are associated withabnormal microglial activation, which appears to be another commonpathology of FTD and ALS (Haukedal et al., (2019) J Mol Biol 431:1818).Other evidence also suggests that ALS and FTD are closely relatedconditions with overlapping genetic, neuropathological, and clinicalfeatures (Weishaupt et al., (2016) Trends Mol Med 22:769; McCauley etal., (2018) Acta Neuropathol 137:715). Taken together, these resultssuggest that both diseases could benefit from shared treatments and thatPGRN genetic variability acts as a modifier of the course of ALS.

In some embodiments, and without wishing to be bound by theory, it isbelieved that anti-Sortilin antibodies of the present disclosure thatinhibit the interaction between Sortilin and Progranulin, neurotrophinsof the present disclosure (e.g., pro-neurotrophins, pro-neurotrophin-3,pro-neurotrophin-4/5, pro-NGF, pro-BDNF, neurotrophin-3,neurotrophin-4/5, NGF, BDNF, etc.), neurotensin, p75, lipoprotein lipase(LpL), apolipoprotein AV (APOA5), and/or receptor associated protein(RAP); or that inhibit one or more activities of Sortilin can beutilized to prevent, or treat one or more undesirable symptoms of ALS

In some embodiments, administering an anti-Sortilin antibody of thepresent disclosure can prevent, reduce the risk, and/or treat ALS. Insome embodiments, administering an anti-Sortilin antibody may modulateone or more Sortilin activities in an individual having ALS.

Depression

As used herein, depression or, major depressive disorder (MDD), clinicaldepression, major depression, unipolar depression, unipolar disorder,recurrent depression or, dysthymia, are used interchangeably and referto a mental disorder characterized by episodes of all-encompassing lowmood accompanied by low self-esteem and loss of interest or pleasure innormally enjoyable activities.

Accordingly, and without wishing to be bound by theory, it is believedthat anti-Sortilin antibodies of the present disclosure that inhibit theinteraction between Sortilin and Progranulin, neurotrophins of thepresent disclosure (e.g., pro-neurotrophins, pro-neurotrophin-3,pro-neurotrophin-4/5, pro-NGF, pro-BDNF, neurotrophin-3,neurotrophin-4/5, NGF, BDNF, etc.), neurotensin, p75, lipoprotein lipase(LpL), apolipoprotein AV (APOA5), and/or receptor associated protein(RAP); or that inhibit one or more activities of Sortilin can beutilized to prevent, or treat one or more undesirable symptoms ofdepression.

In some embodiments, administering an anti-Sortilin antibody of thepresent disclosure can prevent, reduce the risk, and/or treatdepression. In some embodiments, administering an anti-Sortilin antibodymay modulate one or more Sortilin activities in an individual havingdepression.

Parkinson's Disease

Parkinson's disease, which may be referred to as idiopathic or primaryparkinsonism, hypokinetic rigid syndrome (HRS), or paralysis agitans, isa neurodegenerative brain disorder that affects motor system control.The progressive death of dopamine-producing cells in the brain leads tothe major symptoms of Parkinson's. Most often, Parkinson's disease isdiagnosed in people over 50 years of age. Parkinson's disease isidiopathic (having no known cause) in most people. However, geneticfactors also play a role in the disease.

Symptoms of Parkinson's disease include, without limitation, tremors ofthe hands, arms, legs, jaw, and face, muscle rigidity in the limbs andtrunk, slowness of movement (bradykinesia), postural instability,difficulty walking, neuropsychiatric problems, changes in speech orbehavior, depression, anxiety, pain, psychosis, dementia,hallucinations, and sleep problems.

In some embodiments, administering an anti-Sortilin antibody of thepresent disclosure can prevent, reduce the risk, and/or treatParkinson's disease. In some embodiments, administering an anti-Sortilinantibody may induce one or more Progranulin activities in an individualhaving Parkinson's disease. In some embodiments, administering ananti-Sortilin antibody may modulate one or more Sortilin activities inan individual having Parkinson's disease.

Huntington's Disease

Huntington's disease (HD) is an inherited neurodegenerative diseasecaused by an autosomal dominant mutation in the Huntingtin gene (HTT).Expansion of a cytokine-adenine-guanine (CAG) triplet repeat within theHuntingtin gene results in production of a mutant form of the Huntingtinprotein (Htt) encoded by the gene. This mutant Huntingtin protein (mHtt)is toxic and contributes to neuronal death. Symptoms of Huntington'sdisease most commonly appear between the ages of 35 and 44, althoughthey can appear at any age.

Symptoms of Huntington's disease, include, without limitation, motorcontrol problems, jerky, random movements (chorea), abnormal eyemovements, impaired balance, seizures, difficulty chewing, difficultyswallowing, cognitive problems, altered speech, memory deficits,thinking difficulties, insomnia, fatigue, dementia, changes inpersonality, depression, anxiety, and compulsive behavior.

In some embodiments, administering an anti-Sortilin antibody of thepresent disclosure can prevent, reduce the risk, and/or treatHuntington's disease. In some embodiments, administering ananti-Sortilin antibody may induce one or more Progranulin activities inan individual having Huntington's disease. In some embodiments,administering an anti-Sortilin antibody may modulate one or moreSortilin activities in an individual having Huntington's disease.

Taupathy Disease

Taupathy diseases, or Tauopathies, are a class of neurodegenerativedisease caused by aggregation of the microtubule-associated protein tauwithin the brain. Alzheimer's disease (AD) is the most well-knowntaupathy disease, and involves an accumulation of tau protein withinneurons in the form of insoluble neurofibrillary tangles (NFTs). Othertaupathy diseases and disorders include progressive supranuclear palsy,dementia pugilistica (chromic traumatic encephalopathy), frontotemporaldementia and parkinsonism linked to chromosome 17, Lytico-Bodig disease(Parkinson-dementia complex of Guam), Tangle-predominant dementia,Ganglioglioma and gangliocytoma, Meningioangiomatosis, Subacutesclerosing panencephalitis, lead encephalopathy, tuberous sclerosis,Hallervorden-Spatz disease, lipofuscinosis, Pick's disease, corticobasaldegeneration, Argyrophilic grain disease (AGD), Huntington's disease,and frontotemporal lobar degeneration.

In some embodiments, administering an anti-Sortilin antibody of thepresent disclosure, can prevent, reduce the risk, and/or treat taupathydisease. In some embodiments, administering an anti-Sortilin antibodymay induce one or more Progranulin activities in an individual having ataupathy disease. In some embodiments, administering an anti-Sortilinantibody may modulate one or more Sortilin activities in an individualhaving a taupathy disease.

Multiple Sclerosis

Multiple sclerosis (MS) can also be referred to as disseminatedsclerosis or encephalomyelitis disseminata. MS is an inflammatorydisease in which the fatty myelin sheaths around the axons of the brainand spinal cord are damaged, leading to demyelination and scarring aswell as a broad spectrum of signs and symptoms.

Symptoms of MS include, without limitation, changes in sensation, suchas loss of sensitivity or tingling; pricking or numbness, such ashypoesthesia and paresthesia; muscle weakness; clonus; muscle spasms;difficulty in moving; difficulties with coordination and balance, suchas ataxia; problems in speech, such as dysarthria, or in swallowing,such as dysphagia; visual problems, such as nystagmus, optic neuritisincluding phosphenes, and diplopia; fatigue; acute or chronic pain; andbladder and bowel difficulties; cognitive impairment of varying degrees;emotional symptoms of depression or unstable mood; Uhthoffs phenomenon,which is an exacerbation of extant symptoms due to an exposure to higherthan usual ambient temperatures; and Lhermitte's sign, which is anelectrical sensation that runs down the back when bending the neck.

In some embodiments, administering an anti-Sortilin antibody of thepresent disclosure can prevent, reduce the risk, and/or treat multiplesclerosis. In some embodiments, administering an anti-Sortilin antibodymay induce one or more Progranulin activities in an individual havingmultiple sclerosis. In some embodiments, administering an anti-Sortilinantibody may modulate one or more Sortilin activities in an individualhaving multiple sclerosis.

Glaucoma and Macular Degeneration

Glaucoma describes, without limitation, a group of diseases that arecharacterized by a damaged optic nerve, resulting in vision loss andblindness. Glaucoma is usually caused by increased fluid pressure(=intraocular pressure) in the anterior chamber underneath the cornea.Glaucoma results in the successive loss of retinal ganglion cells thatare important for vision. Age-related macular degeneration usuallyaffects older people and primarily causes loss of vision in the macula,the central field of vision. Macular degeneration causes, withoutlimitation, drusen, pigmentary changes, distorted vision, hemorrhages ofthe eye, atrophy, reduced visual acuity, blurred vision, centralscotomas, reduced color vision and reduced contrast sensitivity.

Without wishing to be bound by theory, it is believed that administeringan anti-Sortilin antibody of the present disclosure can prevent, reducethe risk, and/or treat glaucoma and macular degeneration. In someembodiments, administering an anti-Sortilin antibody may induce one ormore Progranulin activities in an individual having glaucoma or maculardegeneration. In some embodiments, administering an anti-Sortilinantibody may modulate one or more Sortilin activities in an individualhaving glaucoma or macular degeneration.

Degenerative Disc Disease (DDD)

Degenerative disc disease (DDD) describes, without limitation, a groupof diseases in which intervertebral disc (IVD) undergoes extensivemorphological as well as biomechanical changes, and usually manifestsclinically in patients with lower back pain. Degenerative discstypically show degenerative fibrocartilage and clusters of chondrocytes,suggestive of repair. Inflammation may or may not be present. Thepathologic findings in DDD include protrusion, spondylolysis, and/orsubluxation of vertebrae (sponylolisthesis) and spinal stenosis.

Without wishing to be bound by theory, it is believed that administeringan anti-Sortilin antibody of the present disclosure can prevent, reducethe risk, and/or treat DDD. In some embodiments, administering ananti-Sortilin antibody may induce one or more Progranulin activities inan individual having DDD. In some embodiments, administering ananti-Sortilin antibody may modulate one or more Sortilin activities inan individual having DDD.

Pain

Pain describes, without limitation, neuropathic pain arising from nerveinjury, e.g., from trauma or disease. Such injury may include injury toperipheral nerves and/or the spinal cord. Sortilin antagonists have beenshown to alleviate neuropathic pain. (Richner M et al. (2019) Sci. Adv.5: eaav9946.)

Kits/Articles of Manufacture

Provided herein are articles of manufacture (e.g., kit) comprising ananti-Sortilin antibody described herein. Article of manufacture mayinclude one or more containers comprising an antibody described herein.Containers may be any suitable packaging including, but is not limitedto, vials, bottles, jars, flexible packaging (e.g., sealed Mylar orplastic bags), and the like. The containers may be unit doses, bulkpackages (e.g., multi-dose packages) or sub-unit doses.

In some embodiments, the kits may further include a second agent. Insome embodiments, the second agent is a pharmaceutically-acceptablebuffer or diluting agent including, but not limited to, such asbacteriostatic water for injection (BWFI), phosphate-buffered saline,Ringer's solution and dextrose solution. In some embodiments, the secondagent is a pharmaceutically active agent.

In some embodiments of any of the articles of manufacture, the articleof manufactures further include instructions for use in accordance withthe methods of this disclosure. The instructions generally includeinformation as to dosage, dosing schedule, and route of administrationfor the intended treatment. In some embodiments, these instructionscomprise a description of administration of the isolated antibody of thepresent disclosure (e.g., an anti-Sortilin antibody described herein) toprevent, reduce risk, or treat an individual having a disease, disorder,or injury selected from dementia, frontotemporal dementia, Alzheimer'sdisease, gauche's disease, vascular dementia, seizures, retinaldystrophy, a traumatic brain injury, a spinal cord injury,atherosclerotic vascular diseases, undesirable symptoms of normal aging,amyotrophic lateral sclerosis (ALS), long-term depression, Parkinson'sdisease, Huntington's disease, Taupathy disease, multiple sclerosis, agerelated macular degeneration, glaucoma, degenerative disc disease (DDD),Creutzfeldt-Jakob disease, normal pressure hydrocephalus, Nasu-Hakoladisease, stroke, acute trauma, chronic trauma, lupus, acute and chroniccolitis, Crohn's disease, inflammatory bowel disease, ulcerativecolitis, malaria, essential tremor, central nervous system lupus,Behcet's disease, mixed dementia, dementia with Lewy bodies, multiplesystem atrophy, Shy-Drager syndrome, progressive supranuclear palsy,cortical basal ganglionic degeneration, acute disseminatedencephalomyelitis, granulomatous disorders, sarcoidosis, diseases ofaging, retinitis pigmentosa, retinal degeneration, respiratory tractinfection, sepsis, eye infection, systemic infection, lupus, arthritis,and wound healing, according to any methods of this disclosure. In someembodiments, the disease, disorder, or injury is frontotemporaldementia. In some embodiments, the instructions include instructions foruse of the anti-Sortilin antibody and the second agent (e.g., secondpharmaceutically active agent).

Diagnostic Uses

The isolated antibodies of the present disclosure (e.g., ananti-Sortilin antibody described herein) also have diagnostic utility.This disclosure therefore provides for methods of using the antibodiesof this disclosure, or functional fragments thereof, for diagnosticpurposes, such as the detection of a Sortilin protein in an individualor in tissue samples derived from an individual.

In some embodiments, the individual is a human. In some embodiments, theindividual is a human patient suffering from, or at risk for developinga disease, disorder, or injury of the present disclosure. In someembodiments, the diagnostic methods involve detecting a Sortilin proteinin a biological sample, such as a biopsy specimen, a tissue, or a cell.An anti-Sortilin antibody described herein is contacted with thebiological sample and antigen-bound antibody is detected. For example, abiopsy specimen may be stained with an anti-Sortilin antibody describedherein in order to detect and/or quantify disease-associated cells. Thedetection method may involve quantification of the antigen-boundantibody. Antibody detection in biological samples may occur with anymethod known in the art, including immunofluorescence microscopy,immunocytochemistry, immunohistochemistry, ELISA, FACS analysis,immunoprecipitation, or micro-positron emission tomography. In certainembodiments, the antibody is radiolabeled, for example with ¹⁸F andsubsequently detected utilizing micro-positron emission tomographyanalysis. Antibody-binding may also be quantified in a patient bynon-invasive techniques such as positron emission tomography (PET),X-ray computed tomography, single-photon emission computed tomography(SPECT), computed tomography (CT), and computed axial tomography (CAT).

In other embodiments, an isolated antibody of the present disclosure(e.g., an anti-Sortilin antibody described herein) may be used to detectand/or quantify, for example, microglia in a brain specimen taken from apreclinical disease model (e.g., a non-human disease model). As such, anisolated antibody of the present disclosure (e.g., an anti-Sortilinantibody described herein) may be useful in evaluating therapeuticresponse after treatment in a model for a nervous system disease orinjury such as frontotemporal dementia, Alzheimer's disease, vasculardementia, seizures, retinal dystrophy, atherosclerotic vasculardiseases, Nasu-Hakola disease, or multiple sclerosis, as compared to acontrol.

The present disclosure will be more fully understood by reference to thefollowing Examples. They should not, however, be construed as limitingthe scope of the present disclosure. All citations throughout thedisclosure are hereby expressly incorporated by reference.

EXAMPLES

Examples 1-3 describe the generation of affinity-matured variants of ananti-human Sortilin antibody, S-60, and the characterization of thosevariants with respect to not only their binding affinity for SORT1 butalso their biological activity in down-regulating SORT1 expression,increasing PGRN secretion, and blocking PGRN-SORT1 binding.Surprisingly, it was found that improved biological activity did notnecessarily correlate with increased affinity.

Example 1: Generation of S-60 Variants and Measurement of SORT1 BindingAffinity

The purpose of the following Example was to generate affinity-maturedvariants of an anti-human Sortilin antibody, S-60, and to characterizethe binding of the affinity-matured antibodies to human Sortilin(SORT1).

S-60 Affinity Matured Variants

Affinity-matured antibodies against human Sortilin (SORT1) weregenerated and their binding to SORT1 measured. The anti-SORT1 antibody,S-60, as well as S-60-1, S-60-2, S-60-3, S-60-4, S-60-5, S-60-6, S-60-7,S-60-8, and S-60-9 have been described in WO2016164637. The V_(H) andV_(L) sequences for S-60-9 and S60 in WO2016164637 are identical, andS-60 will be used herein to refer to the antibody. The V_(H) and VLsequences for S-60-5, S-60-6, and S-60-7 in WO2016164637 are identical,and S-60-7 will be used herein to refer to the antibody. S-60 was foundto be both a potent downregulator of SORT1 levels and a potent blockerof binding between SORT1 and PGRN (see, e.g., Tables 8-10 and FIGS. 6Aand 10D in WO2016164637). Therefore, S-60 was selected for furtheraffinity maturation.

The anti-SORT1 variants S-60-10, S-60-11, S-60-12, S-60-13, S-60-14,S-60-15, S-60-16, S-60-18, S-60-19, and S-60-24 were generated asdescribed below.

Production of S-60 Affinity Matured Variants

The anti-SORT1 antibody S-60 was affinity-matured. Briefly, diversifiedantibody libraries were created in yeast for each of the startingparental antibodies. The diversity in the first round of affinitymaturation was created by utilizing standard molecular cloningtechniques to combine the parental heavy chain HVR-H3 and light chain(LC) with pre-existing genetic diversity in the HVR-H1 and HVR-H2regions of the heavy chain (HC) (termed “H1/H2” optimization). For thesecond round of affinity maturation, both heavy chain variable region(V_(H)) and light chain variable region (VL) sequences were optimized,with a particular focus on HVR-H3.

Selection pressures used for screening the libraries included humanSORT1 antigen equilibrium titration, parental antibody Fab competitionkinetics, and the use of polyspecificity reagent deselection. FACS flowcytometry was then employed to visualize and select antibodies, usingstandard techniques (see, e.g., Chao et al. Nature Protocols, 2006). Thedesired population was then carried forward into additional selectionrounds.

The affinity-matured anti-SORT1 antibodies were purified as follows:clones were grown to saturation and then induced for 48 h at 30° C. withshaking. After induction, yeast cells were pelleted and the supernatantswere harvested for purification. Immunoglobulins were purified using aProtein A column and eluted with acetic acid, pH 2.0. Fab fragments weregenerated by papain digestion and purified over CaptureSelect IgG-CH1affinity matrix (LifeTechnologies).

Measurement of Binding Affinity to SORT1

The anti-SORT1 antibody S-60 and affinity-matured variants thereof weretested and compared in a SORT1 binding assay. For the binding assay,stable HEK293T cells expressing human SORT1 were harvested bytrypsinization, washed in PBS, counted and plated on 96-well u-bottomplates at 1×10⁵ cells/well. The plates were spun at 1,400 rpm for 3minutes and primary anti-SORT1 or control antibodies were added in FACSbuffer (PBS+2 FBS) and incubated on ice for one hour. Cells weresubsequently centrifuged as before and washed thrice with FACS buffer.Cells were then incubated with anti-human APC conjugated secondaryantibody (BD Biosciences) in FACS buffer for 30 minutes on ice. Cellswere again washed three times with FACS buffer and analyzed on a BD FACSCanto or an Intellicyt Flow Cytometer. Binding was measured as medianfluorescent intensity (MFI) in the PE channel of the GFP positive cellpopulation.

Results

The binding affinities of the anti-SORT1 antibodies to SORT1 are shownbelow in Table 1. All antibodies were tested on the huIgG1 backbone.

TABLE 1 Binding Affinity to SORT1. Antibody ID K_(D) (M) Fold changefrom S-60 S-60  1.15E−09* S-60-1 7.221E−09  0.16 S-60-2 1.101E−08  0.10S-60-3  8.56E−10* 1.34 S-60-4 6.34E−10 1.81 S-60-7 7.87E−10 1.46 S-60-84.68E−10 2.46 S-60-10  6.5E−10 1.77 S-60-11  6.5E−10 1.77 S-60-12 8.9E−10 1.29 S-60-13  7.5E−10 1.53 S-60-14 1.05E−09 1.10 S-60-155.60E−10 2.05 S-60-16 7.80E−10 1.47 S-60-18 1.63E−09 0.71 S-60-198.00E−10 1.44 S-60-24  7.55E−10* 1.52 *Note: K_(D) values represent theaverage of two experiments.

Anti-SORT1 antibodies S-60-3, S-60-4, S-60-7, S-60-8, S-60-10, S-60-11,S-60-13, S-60-15, S-60-16, S-60-19, and S-60-24 showed significantlyimproved affinity for binding to SORT1 compared to S-60. In particular,S-60-8 and S-60-15 demonstrated the highest binding affinities to SORT1of the tested variants, having a 2.46- and 2.05-increased fold change,respectively, compared to the binding affinity of the parental antibody,S-60.

Example 2: Effect of S-60 Variants on SORT1 Expression and PGRNSecretion

The purpose of the following Example was to characterize the effect ofaffinity-matured variants of an anti-human SORT1 antibody, S-60, onSORT1 expression and extracellular levels of Progranulin (PGRN).

SORT1 Expression and PGRN Secretion Assays

Affinity matured variants of S-60 were generated as described in Example1 and were subsequently screened by FACS for their abilities todownregulate SORT1 and to elevate extracellular levels of PGRN. In orderto assay the effect of the anti-SORT1 antibodies on SORT1 expression andPGRN secretion, U251 cells that endogenously express SORT1 and secretePGRN were incubated with anti-SORT1 antibody S-60 and affinity-maturedvariants. Cell surface levels of SORT1 at different antibodyconcentrations were measured by FACS as follows: anti-SORT1 antibodyS-60 and affinity-matured variants were added to U251 cells seeded at3×10³ cells/well in 96 well plates with a serial dilution from 0.023 nMto 150 nM. After 72 h, cells were harvested with Trypsin, washed in PBSand labeled with Dylight-650 conjugated anti-SORT1 antibody S-2-11(described in WO2016164637) to quantify the levels of SORT1 proteinremaining. After cells were incubated with 5 μg/ml S-2-11 conjugated toDyLight-650 (Invitrogen) for one hour on ice, cells were washed threetimes in PBS+2 FBS and binding was quantified using a FACSCanto™ orIntellicyt Flow cytometer as median fluorescence intensity (MFI) of APC.

The amount of PGRN secreted into the cell supernatant over the course ofthe 72h was measured by standard ELISA as follows: cells were harvestedand the media was collected; the concentration of PGRN in the mediasamples was measured using an R&D Systems human PGRN Duoset ELISA kit,according to manufacturer's instructions. Data were analyzed inMicrosoft Excel and GraphPad Prism.

Results

The half maximal effective concentration (EC₅₀) for down-regulation (DR)of cell surface levels of SORT1 for each of the tested S-60 variants isshown in Table 2. SORT1 DR was quantified for each S-60 variant testedand expressed as a percent of the control (untreated cells) (Tables2-4). Additionally, the level of extracellular PGRN secretion wasquantified and expressed as a fold change relative to the untreatedcontrol as shown in Tables 2-4. Independent experiments are representedin separate tables (Tables 2-4). All antibodies were tested on thehuIgG1 WT backbone.

TABLE 2 Percent SORT1 Downregulation (DR) and PGRN Secretion for S-60,S-60-1, S-60-2, S-60-3, S-60-4, S-60-7, and S-60-8 PGRN secretion (foldover control) DR at 0.6 nM 0.62 or 0.63 nM Antibody ID DR EC₅₀ (M)antibody (IgG) antibody (IgG) S-60 6.81E−10 29.3 1.38 S-60-1 2.73E−1050.4 1.60 S-60-2 4.20E−10 40.3 1.75 S-60-3 2.69E−10 43.0 1.77 S-60-44.18E−10 39.6 1.61 S-60-7 4.47E−10 32.6 1.40 S-60-8 5.76E−10 32.0 1.54

TABLE 3 Percent SORT1 Downregulation (DR) and PGRN Secretion for S- 60,S-60-3, S-60-24, S-60-15, S-60-16, S-60-18, and S-60-19. PGRN secretion(fold over control), DR at 1.25 nM 0.63 nM Antibody ID antibody (IgG)antibody (IgG) S-60 40.17 1.11 S-60-3 61.50 1.32 S-60-24 65.20 1.35S-60-15 60.92 1.42 S-60-16 70.14 1.42 S-60-18 51.33 1.13 S-60-19 55.461.35

TABLE 4 Percent SORT1 Downregulation (DR) and PGRN Secretion forS-60-24, S-60-10, S-60-11, S-60-12, S-60-13, and S-60-14. PGRN secretion(fold over control), DR at 0.63 nM 0.63 nM Antibody ID antibody (IgG)antibody (IgG) S-60-24 81.48 1.62 S-60-10 85.41 1.85 S-60-11 88.76 1.89S-60-12 87.38 1.89 S-60-13 82.33 1.77 S-60-14 80.47 1.81

In a first experiment analyzing the SORT1 DR EC₅₀, the percent of SORT1DR using a saturating antibody concentration of 0.6 nM IgG, and PGRNsecretion using a saturating antibody concentration of 0.62 or 0.63 nMIgG, the following variants were tested: S-60, S-60-1, S-60-2, S-60-3,S-60-4, S-60-7, and S-60-8. As shown in Table 2, S-60-3 demonstrated thelowest DR EC₅₀ (0.269 nM) and the highest PGRN secretion, a 1.77-foldincrease relative to the control, and an improvement over S-60, whichshowed a 1.38-fold increase over the control. Using 0.6 nM IgG, S-60-1exhibited the most substantial effect on SORT1 expression of thevariants tested, down-regulating SORT1 by 50.4% compared to the control,whereas S-60 down-regulated SORT1 by 29.3%.

In a subsequent experiment, the percent of DR using a saturatingantibody concentration of 1.25 nM IgG, and PGRN secretion using asaturating antibody concentration of 0.63 nM IgG was analyzed for thefollowing variants: S-60, S-60-3, S-60-24, S-60-15, S-60-16, S-60-18,and S-60-19. As shown in Table 3, both S-60-15 and S-60-16 demonstratedthe highest levels of extracellular PGRN secretion with a 1.42-foldincrease compared to the control, an improvement over S-60 which showeda 1.11-fold increase over the control. Indeed, these two variants alsosignificantly reduced SORT1 expression compared to S-60, down-regulatingSORT1 by 60.92% and 70.14%, respectively, relative to the control,whereas S-60 downregulated SORT1 by only 40.17%.

In a third experiment, the percent of DR using a saturating antibodyconcentration of 0.63 nM IgG, and PGRN secretion using a saturatingantibody concentration of 0.63 nM IgG was analyzed for the followingvariants: S-60-24, S-60-10, S-60-11, S-60-12, S-60-13, and S-60-14. Asshown in Table 4, S-60-11 and S-60-12 displayed the highest amount ofextracellular PGRN secretion with a 1.89-fold increase compared to thecontrol. S-60-11 also demonstrated the highest SORT1 DR at 0.63 nM IgG,down-regulating SORT1 by 88.76% relative to the control.

Conclusion

As shown in Table 1 in Example 1, S-60-8 and S-60-15 demonstrated thehighest affinities for binding to SORT1 compared to S-60 and the otherS-60 variants tested with the lowest K_(D) values of 4.68E-10M and5.60E-10, respectively. However, when compared to a specific variant,S-60-3, having a higher K_(D) value of 8.56E-10M (Table 1), S-60-8displayed a smaller increase in PGRN secretion (Table 2) while S-60-15displayed a higher fold increase in PGRN secretion than did S-60-3(Table 3). Additionally, S-60-8 (at a saturating concentration of 0.6 nMIgG) demonstrated a smaller percent reduction of SORT1 expression (Table2) compared to S-60-3 and S-60-15 (at a saturating concentration of 1.25nM IgG), which down-regulated SORT1 by 61.50% and 60.92%, respectively(Table 3). Thus, S-60-15 performed better than S-60-3 with respect toincreasing PGRN secretion and exhibited a similar effect compared toS-60-3 in down-regulating cell surface levels of SORT1, whereas S-60-8performed worse than S-60-3 with respect to increasing PGRN secretionand down-regulating cell surface levels of SORT1.

The results of this study indicate that the relative efficacy ofanti-SORT1 S-60 antibody variants with respect to down-regulating SORT1expression or increasing PGRN secretion was not predicted by theirrelative binding affinities for Sortilin.

Furthermore, S-60-15 showed the greatest increase in PGRN secretioncompared to S-60 and certain S-60 variants (Table 3, last column),including S-60-3. S-60-3, in turn, showed increased PGRN secretionrelative to S-60-1, -2, -4, -7, and -8 (Table 2, last column). ThereforeS-60-15 is an effective antibody for increasing PGRN secretion relativeto those S-60 variants. The ability to increase PGRN secretion is animportant property in selecting a therapeutic antibody for treatment ofdiseases such as FTD and other pathologies in which PGRN deficiencyplays a direct causal role. Such an antibody would be useful inrestoring PGRN levels to, or closer to, wild-type levels, therebytreating the disease. Moreover, this property is an importantpharmacodynamic parameter in assessing antibody activity in vivo (seebelow, Example 5).

Example 3: S-60 Variants Block Binding of PGRN to SORT1

The purpose of the following Example was to characterize the effect ofaffinity-matured variants of an anti-human SORT1 antibody, S-60, on thebinding interaction between SORT1 and a natural ligand, PGRN.

SORT1-PGRN Blocking Assay

The ability of S-60 variants to block the binding of PGRN to SORT1 wasalso tested. Recombinant human PGRN (PGRN) (Adipogen) was biotinylatedwith an EZ-Link Micro NHS-PEG4 kit from ThermoScientific/Pierceaccording to the manufacturer's instructions. A stable cell lineexpressing full-length untagged human SORT1 (SORT1) was established byviral infection of HEK293T cells, and positive selection with hygromycin(Genscript custom project). As control cells, parental HEK293T cellswere utilized.

SORT1-expressing cells or control cells were harvested and washed inPBS. Biotinylated human PGRN was added in PBS+2% FBS with or without atitration of anti-SORT1 antibodies or control human IgG1 isotypeantibodies and incubated on ice for 2h. After washing cells 3 times inPBS+2% FBS, cells were incubated in Streptavidin-APC (BD Biosciences,1:100) on ice for 30 min. Then cells were washed again, resuspended inPBS+2% FBS and analyzed on a FACSCanto™ flow cytometer (BD Biosciences,Mississauga, ON). PGRN binding was measured as the median fluorescenceintensity (MFI) of APC of the SORT1 expressing cell population. Bindingof biotinylated PGRN to SORT1 expressed on HEK293T cells was measured byFACS in absence or presence of S-60 and its variant antibodies.

Results

The half maximal effective concentration (EC₅₀) for blocking PGRNbinding to SORT1 for each of the tested S-60 variants is shown in Tables5-7. Additionally, the maximum level of blocking PGRN to SORT1 wasquantified and expressed as a percentage of the maximum block achievedusing a saturating concentration of 50 nM IgG or 150 nM IgG (Table 5-7).Independent experiments are shown in separate tables below.

TABLE 5 Blocking of PGRN binding to SORT1 expressed on HEK293T cells forS-60-1, S-60-2, S-60-3, S-60-4, S-60-7, S-60-8, and S-60, PGRN bindingblock PGRN binding block (% max at 150 nM Antibody ID EC₅₀ (nM) antibody(IgG)) S-60-1 5.986 95.2 S-60-2 7.509 94.5 S-60-3 0.733 95.5 S-60-40.584 95.7 S-60-7 0.944 96.5 S-60-8 0.985 95.6 S-60 0.751 95.9

TABLE 6 Blocking of PGRN binding to SORT1 expressed on HEK293T cells forS-60, S-60-3, S-60-24, S-60-15, S-60-16, S-60-18, and S-60-19. PGRNbinding block PGRN binding block (% max at 50 nM Antibody ID EC₅₀ (M)antibody (IgG)) S-60 2.20E−09 90.13 S-60-3 1.76E−09 90.44 S-60-241.11E−09 88.94 S-60-15 1.22E−09 90.74 S-60-16 1.20E−09 90.44 S-60-182.27E−09 89.33 S-60-19 1.53E−09 90.15

TABLE 7 Blocking of PGRN binding to SORT1 expressed on HEK293T cells forS-60-24, S-60-10, S-60-11, S-60-12, S-60-13, and S-60-14. PGRN bindingblock PGRN binding block (% max at 50 nM Antibody ID EC₅₀ (M) antibody(IgG)) S-60-24 4.70E−10 95.64 S-60-10 4.60E−10 93.84 S-60-11 4.10E−1094.97 S-60-12 6.10E−10 93.26 S-60-13 5.60E−10 93.83 S-60-14 6.00E−1093.67

In a first experiment analyzing the PGRN binding block EC₅₀ and thepercentage of PGRN binding block to SORT1 with a saturatingconcentration of 150 nM IgG, the following variants were tested: S-60-1,S-60-2, S-60-3, S-60-4, S-60-7, S-60-8, and S-60. As shown in Table 5,S-60-4 demonstrated the lowest EC₅₀ value (0.584 nM), achieving a 95.7%block of PGRN binding to SORT1. Of the variants analyzed, S-60-7displayed the greatest reduction in the PGRN binding, blocking 96.5% ofPGRN binding to SORT1, though it displayed lower efficiency at blockingPGRN binding (higher EC₅₀ value) than S-60 and variants S-60-3 andS-60-4.

In a subsequent experiment analyzing the PGRN binding block EC₅₀ and thepercentage of PGRN binding block to SORT1 with a saturatingconcentration of 50 nM IgG, the following variants were tested: S-60,S-60-3, S-60-24, S-60-15, S-60-16, S-60-18, and S-60-19. As shown inTable 6, S-60-24, S-60-15, and S-60-16 displayed the lowest EC₅₀ valuescompared to S-60, ranging from 1.11 nM to 1.22 nM. Furthermore, theS-60-15 variant demonstrated the strongest potency compared to S-60,achieving a 90.74% block of PGRN binding to SORT1.

In a third experiment, the following variants were tested: S-60-24,S-60-10, S-60-11, S-60-12, S-60-13, and S-60-14. As shown in Table 7,S-60-11 demonstrated the lowest EC₅₀ (0.410 nM), achieving a 94.97%block of PGRN binding to SORT1. Of the variants analyzed, S-60-24displayed the greatest reduction in the PGRN binding, blocking 95.64% ofPGRN binding to SORT1.

Conclusion

As shown in Table 1 in Example 1, both S-60-8 and S-60-15 demonstratedthe highest affinities for binding to SORT1 relative to S-60 and theother S-60 variants tested, with S-60-8 having the highest affinity forSORT1. However, when compared to S-60, S-60-8 displayed a lowerefficiency at blocking binding to PGRN (higher EC₅₀ value relative toS-60) (Table 5) whereas S-60-15 displayed a higher efficiency atblocking binding to PGRN (reduced EC₅₀ value relative to S-60) (Table6). Moreover, S-60-8 demonstrated a smaller percent maximal blocking ofPGRN binding to SORT1 compared to S-60 (Table 5) whereas S-60-15demonstrated a greater percent maximal blocking of PGRN binding to SORT1relative to S-60 (Table 6). Thus, S-60-15 performed better than S-60-8when compared to S-60 with respect to EC₅₀ value in blocking PGRNbinding to SORT1 and the maximum percentage of blocking PGRN binding toSORT1, despite the fact that S-60-8 showed higher affinity binding toSORT1 compared to S-60-15.

Thus, the results of this study indicate, unexpectedly, that therelative efficacy of anti-SORT1 S-60 antibody variants with respect toblocking the degree of PGRN binding to SORT1 was not predicted by theirrelative binding affinities for SORT1.

Example 4: S-60-15 Stability and Stress Testing Analysis

The purpose of the following Example was to characterize S-60-15, anaffinity-matured variant of S-60 described in previous Examples, forstability under various stress testing conditions.

Stability of a therapeutic antibody is important for clinical efficacy.During manufacturing, storage, and in vivo administration, therapeuticantibodies are at risk for degradation via multiple pathways. Thefactors that are responsible for such degradation remain poorlyunderstood. Given that S-60-15 demonstrated a high binding affinity forSORT1 (see Example 1), effectively reduced cell surface levels of SORT1and increased extracellular PGRN secretion (see Example 2), as wellsubstantially blocked PGRN binding to SORT1 (see Example 3), andunexpectedly performed these functional tests better than S-60-8 whichshowed higher affinity for SORT1, S-60-15 was further evaluated forstability.

pH and Temperature Stress Testing

In order to measure the stability of the amino acid residues withinS-60-15, samples of S-60-15 were subjected to pH and temperature stressconditions to emulate stress conditions that occur during manufacturing,storage, and in vivo administration. Briefly, individual samples ofS-60-15 were subjected to stress testing at pH 3.5 or pH 5.0 and at 40°C. or 50° C. The samples were tested at various time points—0, 1, 3 and5 days for the two pH conditions, and 0, 7, 14 and 30 days for the hightemperature stress conditions—by treatment with DTT and IAA followed bytrypsin digestion. The digested samples were analyzed by LiquidChromatography with Mass Spectroscopy detection using a Waters ACQUITYUPLC coupled to a Xevo G2-XS QTOF mass spectrometer using a BEH C18column. Results are shown in Table 8.

TABLE 8Stoichiometry Percentage Table (% of each peptide with the modifier). Deamid (deamidation); Oxid (oxidation ; CAM (carbamido methyl).Non- pH5 pH5 pH5 pH5 PH3.5 PH3.5 PH3.5 PH3.5 T40 T40 T40 T50 T50 T50S-60-15 Peptide Modifier stress d0 d1 d3 d5 d0 d1 d3 d5 d7 d14 d30 d7d14 d30 NQFSLK (SEQ ID NO: 87) Deamid  1.3 1.5 1.5 1.5  0.9 1.5 1.4 1.51.4  1.3  1.3  1.3  1.3  1.3  1.4 QGYYGMDVWGQGTTVTVSSAS Oxid M(1)  6.26.8 6.2 6.6  8.8 6.2 6.8 6.6 6.4  5.7  5.1  5.6  5.1  5.4  1.3TK (SEQ ID NO: 88) QGYYGMDVWGQGTTVT Deamid  0.5 0.6 0.6 0.6  0.7 0.6 0.60.6 0.6  0.8  1.0  1.0  1.4  2.3  3.6 VSSASTK (SEQ ID NO: 88) Q(1)DYFPEPVTVSWNSGALTSGV Deamid  3.3 3.3 3.3 3.3  3.8 3.4 3.4 3.4 3.2  3.2 3.1  0.0  3.2  3.1  3.2 HTFPAVLQSSGLYSLSSVVTV N(1)PSSSLGTK (SEQ ID NO: 89) Oxid W(1) DYFPEPVTVSWNSGALTSGV Deamid  0.3 0.40.4 0.4  0.4 0.4 0.4 0.3 0.3  0.4  0.5  0.8  0.7  1.0  1.5HTFPAVLQSSGLYSLSSVVTV N(1) PSSSLGTK (SEQ ID NO: 89)DTLMISR (SEQ ID NO: 90) Oxid M(1)  3.1 3.6 3.4 3.5  3.5 3.6 3.1 3.4 3.3 4.0  4.4  5.8  5.2  7.2 12.2 TPEVTCVVVDVSQEDPEV Deamid  1.4 1.1 1.2 1.3 1.7 1.5 1.2 1.4 1.3  0.2  0.3  1.3  1.1  0.2  5.2 QFNWYVDGVEVHNAK N(1)(SEQ ID NO: 91) VVSVLTVLHQDWLNGK Deamid  7.8 7.2 6.7 6.9  6.5 7.0 6.97.0 6.8  8.1  8.4  9.2  9.3 12.6 12.0 (SEQ ID NO: 92) N(1)EPQVYTLPPSQEEMTK Oxid. M(1)  0.3 0.3 0.0 0.0  1.5 0.2 0.4 0.4 0.5  0.8 0.0  0.0  0.8  1.3  1.1 (SEQ ID NO: 93) NQVSLTCLVK (SEQ ID NO: 94)Deamid  1.9 2.1 2.0 2.0  1.7 2.0 2.0 2.0 1.9  1.9  2.1  1.9  2.1  2.5 2.0 N(1) CAM C(1) GFYPSDIAVEWESNGQPENNYK Deamid  5.2 4.9 5.1 5.0  6.45.0 5.0 5.2 4.9  7.0  8.9  9.6 11.1 19.0 45.9 (SEQ ID NO: 95) N(1)WQEGNVFSCSVMHEALH Deamid  3.8 4.0 4.0 4.0  4.4 3.9 3.0 4.0 3.9  3.7  3.7 3.9  3.9  4.4  6.7 NHYTQK (SEQ ID NO: 96) N(1) CAM C(1)WQEGNVFSCSVMHEALH CAM C(1),  1.6 1.8 1.8 1.8  1.9 1.8 1.8 1.9 1.8  1.8 1.9  2.6  2.2  2.6  5.1 NHYTQK (SEQ ID NO: 96) Oxid. M(1)DIVMTQSPLSLPVTPGEPASISCR CAM C(1),  0.7 0.8 0.8 0.8  0.8 0.8 0.7 0.8 0.7 0.8  0.8  1.1  0.8  0.8  2.0 (SEQ ID NO: 97) Oxid. M(1)SSQSLLR (SEQ ID NO: 98) Deamid  6.8 6.1 5.2 5.4  6.5 6.0 5.4 6.7 5.8 7.3  6.9  7.0  6.8  7.2  6.6 Q(1) SNGYNYLDWYLQKPGQSP Deamid 11.2 9.39.4 9.3 12.4 9.8 9.7 9.8 9.6 15.7 20.3 21.9 30.7 45.4 65.6QLLIYLGSNR (SEQ ID NO: 99) N(1) AEAEDVGVYYCMQQQEAP CAM C(1),  0.5 0.70.7 0.7  0.7 0.7 0.7 0.7 0.7  0.5  0.6  1.1  0.8  1.1  2.1LTFGGGTK (SEQ ID NO: 100) Oxid. M(1) SGTASVVCLLNNFYPR Deamid  2.6 2.82.9 2.8  3.1 2.8 2.7 2.8 2.7  2.6  2.7  2.5  2.6  3.0  3.4(SEQ ID NO: 101) N(1) VDNALQSGNSQESVTEQDSK Deamid  0.3 0.4 0.4 0.4  0.40.3 0.4 0.4 0.4  0.5  0.5  0.5  0.6  0.8  1.3 (SEQ ID NO: 102) N(1)

Amino acids having the potential to incur post-translationalmodifications (PTMs) upon stress were identified, and the correspondingpeptide fragments generated from the stressed samples were trackedkinetically. The results confirm that two asparagine-glycine sites, onein the Fc region of the heavy chain (SEQ ID NO:11) and one in the Fabregion of the light chain (SEQ ID NO:15) of S-60-15, were the mostsusceptible to PTM, exhibiting 45.9% and 65.6% deamidation,respectively, after being subjected to 50° C. storage for 30 days. Noother PTM or cleavage site exhibited notable PTM or degradation uponstress testing.

Deamidation of the Asparagine (N33) Site

Any deamidation in the Fc region of the heavy chain of S-60-15 shouldnot affect the binding affinity of S-60-15 to SORT1, as that region doesnot directly interact with SORT1. However, the asparagine (N33) site inthe Fab region of S-60-15 is in the HVR-L1 (SEQ ID NO:8), thecomplementarity determining region of the light chain, whichparticipates in antigen binding. Thus, deamidation of the asparagine(N33) site in the Fab region of S-60-15 may affect the binding affinityof S-60-15 to SORT1. Upon deamidation, the asparagine (N33) site in theFab region of S-60-15 undergoes an Asn to Asp/IsoAsp change.

Site-directed mutagenesis was utilized to test the effects of thedeamidation of the asparagine (N33) site in the Fab region of S-60-15and the potential amino acid substitution to reduce the likelihood ofdegradation and deamidation and resulting manufacturing liability.Seventeen different point mutations to the HVR-L1 N33 position ofS-60-15 huIgG1 were produced and sequenced using standard procedures.The G1m3 or G1m(f) allotype with L234A/L235A/P331S (“LALAPS” motif)mutations was used as a background (Jefferis R and Lefranc M-P, Mabs,2009 July-August; 1(4): 332-338). The LALAPS point mutations areintended to improve overall safety by minimizing effector functions suchas Fc gamma receptor binding, complement activation, and antibodydependent cell mediated cytotoxicity (ADCC). Reduction of Fc gammareceptor binding, complement activation and ADCC as a result of LALAPSmutations have been observed in various experiments (data not shown).Bio-layer interferometry (BLI) data was collected at a rate of 5 Hz at25° C. on a Pall ForteBio Octet RED96 instrument. Data analysis wasperformed using ForteBio Data Analysis Software, version 9.0. Standardkinetic buffer (PBS, 0.1 BSA, 0.02 Tween-20, pH 7.2) was used for theassay and for preparing reagents. For all assays, sensor tips wereequilibrated in buffer prior to analysis.

For the initial off-rate screen, S-60-15 LALAPS N33 and N33X mutants (1μg/mL, 300 s loading time) were captured on Anti-Human IgG Fc CaptureDip and Read Biosensors (Pall ForteBio, Menlo Park, Calif.). Twenty nMhistidine-tagged human SORT1 (R&D Systems, Minneapolis, Minn.) was thenbound to the captured anti-SORT1 surface (200 s association time, 1200 sdissociation time). The resulting BLI signal was obtained as thedifference in response from the reference (1 μg/mL S60-15+0 nM SORT1)sensor. A zero-ligand control (0 μg/mL IgG+20 nM SORT1) showed nomeasurable non-specific binding of antigen to the sensor tip surface.Full, local fitting using a 1:1 interaction model was applied to extractassociation and dissociation rate constants (k_(a) and k_(d),respectively) for each N33 mutant. Affinity constants (K_(D)) werecalculated from the ratio k_(a)/k_(d). Different amino acidsubstitutions were made at N33 and tested for binding to SORT1 in anoff-rate screen (under non-stress conditions) as shown in Table 9.

TABLE 9 Extracted k_(a,) k_(d), and K_(D) for S-60-15 N33X Mutantsbinding to SORT1 S-60-15 Variants k_(a) (Ms)⁻¹ k_(d) (s⁻¹) K_(D) (pM)S-60-15 [N33 (wt)] 5.12E+05 1.41E−04 270 S-60-15.1 [N33T]  4.7E+051.67E−04 344 S-60-15.2 [N33S] 5.63E+05 2.47E−04 440 S-60-15.3 [N33G]4.98E+05 4.11E−04 825 S-60-15.4 [N33R] 5.24E+05 4.60E−04 880 S-60-15.5[N33D] 5.90E+05 6.10E−04 1000 S-60-15.6 [N33H] 5.48E+05 4.60E−04 823S-60-15.7 [N33K] 5.63E+05 5.47E−04 965 S-60-15.8 [N33Q] 6.09E+056.53E−04 905 S-60-15.9 [N33Y] 5.11E+05 6.75E−04 1285 S-60-15.10 [N33E]6.76E+05 4.30E−03 2007 S-60-15.11 [N33W] 6.01E+05 5.79E−04 960S-60-15.12 [N33F] 2.80E+05 8.16E−04 2910 S-60-15.13 [N33I] 4.91E+052.28E−04 460 S-60-15.14 [N33V] 4.45E+05 3.06E−04 690 S-60-15.15 [N33A]4.93E+05 3.15E−04 625 S-60-15.16 [N33M] 4.95E+05 4.23E−04 850 S-60-15.17[N33L] 4.12E+05 4.25E−04 1030 Note: k_(a,) k_(d), and K_(D) represent anaverage value from multiple experiments.

Screening this N33X mutant panel showed that the S-60-15.5 antibody(N33D), which mimics deamidation of N33, led to a significant reductionin binding, with a K_(D) of 1000 pM compared to 270 pM for N33 wild-type(wt) (Table 9). In contrast, the S-60-15.1 antibody (N33T) had the bestretention of S-60-15 affinity to SORT1, displaying the smallest K_(D)difference from wild-type in the panel (Table 9).

In addition, complete kinetic analysis using multiple concentrations ofSORT1 was performed for S-60-15.1 [N33T] as huIgG1 and huIgG1 LALAPS inan independent experiment. The kinetic data are displayed in Table 10.Kinetic data demonstrate that S60-15.1 [N33T] is a high affinityantibody both as huIgG1 and huIgG1 LALAPS.

TABLE 10: SORT1-binding kinetic data for S-60-15.1 [N33T] as huIgG1 andhuIgG1 LALAPS. Antibody ID k_(a) (Ms)⁻¹ k_(d) (s⁻¹) K_(D) (pM) S-60-15.1[N33T] 5.99E+05 1.79E−04 298 S-60-15.1 [N33T] LALAPS 4.79E+05 1.27E−04260

Cell Binding and Functionality

S-60-15.1 [N33T], both as huIgG1 and huIgG1 LALAPS, and S-60-15.5 [N33D]were subsequently tested for cell binding and functionality as describedbelow.

Affinity matured antibodies S-60, S-60-15.1 [N33T] and S-60-15.5 [N33D]were tested side-by-side for binding to human SORT1 expressed on HEK293Tcells as described earlier. The results are shown in Table 11. S-60-15.1[N33T] as huIgG1 or huIgG1 LALAPS showed a higher affinity than S-60(Table 11), with a lower K_(d) and a higher B_(max). The Fc variant didnot appear to have an effect on affinity, having only marginaldifferences in K_(d) and B_(max) of S-60-15.1 [N33T] huIgG1 vs. huIgG1LALAPS. The completely deamidated version of S-60-15.5 [N33D] did notshow any significant binding (Table 11, N.B. (No Binding)) to humanSORT1 expressing cells, suggesting that amidation of the N33 residue isessential for target binding.

TABLE 11 Cell binding affinity values as measured by FACS. Antibody IDK_(d) (nM) B_(max) (MFI) S60 1.02 758381 S-60-15.5 [N33D] N.B. N.B.S-60-15.1 [N33T] 0.3651 791996 S-60-15.1 [N33T] LALAPS 0.4231 792791

U251 cells that endogenously express SORT1 and secrete PGRN wereincubated with anti-SORT1 antibody S-60, S-60-15.1 [N33T] or S-60-15.5[N33D] at different concentrations, and cell surface levels of SORT1were measured by FACS as described in Example 2. The control wasuntreated cells. In order to quantify the percentage of Sortilindown-regulation, a saturating antibody concentration of 150 nM IgG wasused. For quantification of PGRN secretion, a saturating antibodyconcentration of 50 nM IgG was utilized. Results are shown in Table 12.S-60-15.1 [N33T] as huIgG1 or huIgG1 LALAPS resulted in significantdownregulation of SORT1 and an increase in PGRN secretion. Both thelevel of downregulation of SORT1 and the level of increase in PGRNsecretion were significantly improved in 5-60-15.1 [N33T] as huIgG1 orhuIgG1 LALAPS as compared to S-60. S-60-15.1 [N33T] as huIgG1 displayedonly a minor reduction in EC₅₀ and percent SORT1 down-regulation, havinga minor fold increase in PGRN secretion compared to S-60-15.1 [N33T] ashuIgG1 LALAPS. In contrast, the completely deamidated version ofS-60-15.5 [N33D] performed worse than S-60 in each of these functionalassays; it did not show significant downregulation of SORT1 and had onlyminimal change in secreted PGRN levels.

TABLE 12 Downregulation (DR) of cell surface SORT1 by S60 antibodyvariants. PGRN secretion, fold % DR at 150 nM over control at 50 nMAntibody ID EC₅₀ (pM) antibody (IgG) antibody (IgG) S-60 680.9 62.5 1.75S-60-15.5 [N33D] n.d. 3.4 1.22 S-60-15.1 [N33T] 72.58 69.3 2.29S-60-15.1 [N33T] 103.6 70.3 1.97 LALAPS

Binding of biotinylated PGRN to SORT1 expressed on HEK293T cells wasmeasured by FACS in the absence or presence of anti-SORT1 antibody S-60,S-60-15.1 [N33T] and S-60-15.5 [N33D] as described in Example 3. Resultsare shown in Table 13. A saturating antibody concentration of 150 nM IgGwas used for quantifying the percentage of blocking of PGRN binding toSORT1.

TABLE 13 Block of PGRN binding to SORT1 expressed on HEK293T cells.Percentage block at 150 nM Antibody ID EC₅₀ (nM) antibody (IgG) S600.751 95.9 S-60-15.5 [N33D] 0.588 27.5 S-60-15.1 [N33T] 0.325 96.5S-60-15.1 [N33T] LALAPS 0.405 96.9

The S-60-15.1 [N33T] variants, both with and without LALAPS, displayedthe lowest EC₅₀ values and the highest level of blocking PGRN binding toSORT1 as compared to S-60-15.5 as well as compared to S-60. Thus,S-60-15.1 [N33T] performed better than S-60 and S-60-15.5 in blockingPGRN binding to SORT1.

In summary, S-60-15 was found to undergo extensive deamidation at N33under storage stress (see Table 8), which increases the likelihood ofdegradation and resulting manufacturing liability. The previouslydescribed anti-SORT1 antibody S-60 and variants S-60-1, S-60-2, S-60-3,S-60-4, S-60-7, and S-60-8 also contain an asparagine at position 33 inthe V_(L). In particular, the location of the N33 deamidation sitewithin the HVR-L1 site is likely to affect the binding of S-60-15 toSORT1. S-60-15.1 [N33T], one of several tested S-60-15 variants,retained significant binding affinity to SORT1 as compared to S-60-15(see Table 9) and showed significantly improved binding affinity toSORT1 as compared to S-60 (see Table 11; compare to S-60-15 WT in Table1). This high retention of binding affinity was unexpected for anantibody having a non-preferred substitution within HVR-L1 because theHVR-L1 residues may contact antigen. Also, unexpectedly, the S-60-15.1[N33T] variant retained increased downregulation of SORT1, increasedsecretion of PGRN, and increased blocking of PGRN binding to SORT1 ascompared to S-60 (see Tables 12 and 13). These results were alsounexpected because a non-preferred substitution in an HVR sequence wouldlikewise have been expected to negatively affect the functionalcharacteristics of an antibody. Instead, S-60-15.1 [N33T] retains thedesirable properties of downregulating cell-surface SORT1, increasingPGRN secretion, and blocking binding of PGRN to SORT1 in addition tobeing more stable than previously described antibodies by removal of amanufacturing liability at position 33 in the VL.

Example 5: Anti-Sortilin Antibody PK and PD in Non-Human Primates

In this Example, the pharmacokinetics (PK) and pharmacodynamics (PD) ofintravenously (IV) administered anti-Sortilin antibody S-60-15.1 [N33T]LALAPS were determined in non-human primates.

Materials and Methods

Single Dose Pharmacokinetic and Pharmacodynamic Studies

For single dose pharmacokinetic studies, cynomolgus monkeys wereadministered anti-Sortilin antibody by single IV dose of 5 mg/kg, 20mg/kg, 60 mg/kg, or 200 mg/kg on Day 0 (n=3 animals per dose). Blood andCSF were drawn from the animals at multiple time-points thereafter toobtain anti-Sortilin antibody concentrations in plasma and cerebrospinalfluid (CSF), which are measurements of anti-Sortilin antibodypharmacokinetics. Progranulin (PGRN) concentration and the levels ofSortilin (SORT1) on white blood cells (WBCs), which are measurements ofpharmacodynamics, were also determined.

Anti-Sortilin antibody concentrations were assayed using an ELISA assaywith anti-Sortilin antibody-specific anti-idiotypic antibodies. PGRNconcentrations were assayed with a commercially-available ELISA kit.Levels of SORT1 on white blood cells were assayed using an ELISA assay,and normalized to protein concentration.

Results

Table 14 provides the plasma mean C_(max), mean AUC, and t_(1/2) foreach of the tested anti-Sortilin antibody doses.

TABLE 14 C_(max), mean AUC, and t_(1/2) for the indicated anti- Sortilinantibody doses (n = 3 for each dose). Mean C_(max) Mean AUC AntibodyDose (μg/ml) (μg × hr/ml) t ½ hours 5 mg/kg 156 2,870 4.7 20 mg/kg 69726,500 13.3 60 mg/kg 2,570 118,000 42 200 mg/kg 7,910 366,000 73.6

As shown in FIG. 1A, SORT1 expression levels in peripheral white bloodcells decreased after treatment of non-human primates with any of theanti-Sortilin antibody doses tested. The higher anti-Sortilin antibodydoses (60 mg/kg, 200 mg/kg) resulted in both an earlier and moreprolonged decrease of SORT1 levels in peripheral white blood cellscompared to lower anti-Sortilin antibody doses (5 mg/kg, 20 mg/kg).

The levels of PGRN increased in the plasma of non-human primatesadministered a single IV injection of anti-Sortilin antibody in a time-and dose-dependent manner (FIG. 1B). In particular, plasma PGRN levelsincreased 3- to 4-fold at C_(max), compared to baseline levels, for allanti-Sortilin antibody doses tested. Plasma PGRN levels remainedelevated for longer periods of time at the higher antibody doses.Additionally, increased plasma PGRN levels were correlated withdecreased expression levels of SORT1 in peripheral white blood cells.

The levels of PGRN in CSF were also increased in non-human primatesadministered a single IV injection of anti-Sortilin antibody. As shownin FIG. 1C, CSF PGRN levels increased 2- to 3-fold above baseline inanimals administered either 20 mg/kg, 60 mg/kg, or 200 mg/kg. Asobserved with plasma PGRN levels, CSF PGRN levels remained elevated overtime in the higher antibody dose groups.

Table 15 provides the CSF mean C_(max), mean AUC, and t_(1/2) for eachof the tested anti-Sortilin antibody doses in non-human primates.Anti-Sortilin antibody CSF concentrations were on average around 0.1%the amount observed in plasma.

TABLE 15 Anti-Sortilin antibody CSF PK parameters and estimatedhalf-life in non-human primates. [1] Dose C_(max) AUC_(all) CL t_(1/2)hours Level (μg/mL) (h*μg/mL) (mL/h/kg) (days) 5 mg/kg 20 184 20692 32.3(1.34) 20 mg/kg 2243 35717 745 23.8 (1) 60 mg/kg 6842 113573 623 38.3(1.6) 200 mg/kg 4595 349187 1037 72.4 (3.02)

Repeat Dose Pharmacokinetic and Pharmacodynamic Studies

Further pharmacokinetic and pharmacodynamic studies were performed innon-human primates administered anti-Sortilin antibody following arepeat-dose regimen. In these studies, animals (2 males and 2 females)were administered anti-Sortilin antibody at a dose of 60 mg/kg once perweek for four weeks. At various timepoints thereafter, SORT1 expressionlevels in peripheral white blood cells were determined. In addition,plasma and CSF levels of the anti-Sortilin antibody, were determined.

As shown in FIG. 2A, SORT1 levels in peripheral white blood cellsremained decreased throughout the duration of the study. Plasma PGRNlevels increased to 5- to 6-fold above baseline at peak levels (FIG.2B). A decrease in plasma PGRN was observed following the fourth andfinal administration of anti-Sortilin antibody; however, the plasma PGRNlevels remained elevated by 2-fold above baseline. Additionally, CSFPGRN levels were increased 3- to 4-fold above baseline (FIG. 2C).

The systemic anti-Sortilin antibody exposure, assessed by mean Cmax andAUC₀₋₁₆₈, was 2100 μg/mL and 114,000 μg/mL×hr on Day 1, and 3020 μg/mLand 174,000 μg/mL×hr on Day 22. These results showed that exposure washigher on Day 22 compared to Day 1, indicating some accumulation of theantibody.

CSF concentration of anti-Sortilin antibody in these animals ranged from0.03% to 0.12% of that observed in plasma, consistent with thedistribution of other antibodies in the CSF (Pestalozzi et al., (2000) JClin Oncol 18(11):2349-51; Petereit et al., (2009) Mult Scler15(2):189-92).

Taken together, these results indicate that the S-60-15.1 [N33T]variant, in spite of having a relatively short half-life, has sustainedactivity in vivo, decreasing SORT1 levels in peripheral white bloodcells and increasing plasma and CSF PGRN levels.

Moreover, IV administration of S-60-15.1 [N33T] to cynomolgous monkeysin both a 4-week and a 26-week repeat dose toxicology study was welltolerated at up to 200 mg/kg weekly, and there were no adverse findingsthat would preclude the conduct of clinical studies in humans based onthose and other toxicology studies which included assessments ofcytokine release, mortality, body weight, respiratory rate and depth,and local tolerability at injection site. Therefore, S-60-15.1 [N33T] issuitable for testing in human clinical studies of FTD and otherindications as contemplated herein.

Lack of adverse findings with respect to toxicity from administration ofan anti-Sortilin antibody was surprising in view of the fact thatSortilin acts as a receptor of several ligands and has multiple roles incellular transport and signaling (Nykjaer, A et al., (2012) TrendsNeurosci 35: 261-270). Due to Sortilin's multiple ligands and functionalroles, administration of an anti-Sortilin antibody might have beenexpected to cause off-target effects; however, the S-60-15.1 [N33T]antibody was well tolerated at the administered doses.

Sequences of the V_(H) regions of the S-60 antibody variants, alignment to S-60, and HVR locationsS-60 (SEQ ID NO: 56)QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGSTYY 60 S-60-1(SEQ ID NO: 107)QVQLQESGPGLVKPSETLSLTCAVSGYSISSVRYWGWIRQPPGKGLEWIGSIYHSGSTYY 60 S-60-2(SEQ ID NO: 108)QVQLQESGPGLVKPSETLSLTCAVSGYSISSVRYWGWIRQPPGKGLEWIGAIYPSGSTYY 60 S-60-3(SEQ ID NO: 109)QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGSTYY 60 S-60-4(SEQ ID NO: 110)QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGSTYY 60 S-60-7(SEQ ID NO: 111)QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGSTYY 60 S-60-8(SEQ ID NO: 112)QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGSTYY 60 S-60-10(SEQ ID NO: 54)QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGSTYY 60 S-60-11(SEQ ID NO: 54)QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGSTYY 60 S-60-12(SEQ ID NO: 54)QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGSTYY 60 S-60-13(SEQ ID NO: 55)QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGSTYY 60 S-60-14(SEQ ID NO: 55)QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGSTYY 60 S-60-15(SEQ ID NO: 56)QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGSTYY 60S-60-15.1 (SEQ ID NO: 56)QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGSTYY 60 S-60-16(SEQ ID NO: 56)QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGSTYY 60 S-60-18(SEQ ID NO: 56)QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGSTYY 60 S-60-19(SEQ ID NO: 54)QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGSTYY 60 S-60-24(SEQ ID NO: 56)QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGWIRQPPGKGLEWIGTIYHSGSTYY 60 S-60Alignment *******************************  *****************:** ******S-60 HVR H1 (SEQ ID NO: 1)--------------------------YSISSGYYWG------------------------ S-60HVR H2 (SEQ ID NO: 2)--------------------------------------------------TIYHSGSTYY S-60(SEQ ID NO: 56 cont.)NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTTVTVSS 122S-60-1 (SEQ ID NO: 107 cont.)NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTTVTVSS 122S-60-2 (SEQ ID NO: 108 cont.)NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTTVTVSS 122S-60-3 (SEQ ID NO: 109 cont.)NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTTVTVSS 122S-60-4 (SEQ ID NO: 110 cont.)NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTTVTVSS 122S-60-7 (SEQ ID NO: 111 cont.)NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTTVTVSS 122S-60-8 (SEQ ID NO: 112 cont.)NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTTVTVSS 122S-60-10 (SEQ ID NO: 54 cont.)NPSLKSQVTISVDTSKNQFSLELSSVTAADTAVYYCARQGSIQQGYYGMDVWGQGTTVTVSS 122S-60-11 (SEQ ID NO: 54 cont.)NPSLKSQVTISVDTSKNQFSLELSSVTAADTAVYYCARQGSIQQGYYGMDVWGQGTTVTVSS 122S-60-12 (SEQ ID NO: 54 cont.)NPSLKSQVTISVDTSKNQFSLELSSVTAADTAVYYCARQGSIQQGYYGMDVWGQGTTVTVSS 122S-60-13 (SEQ ID NO: 55 cont.)NPSLESRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIQQGYYGMDVWGQGTTVTVSS 122S-60-14 (SEQ ID NO: 55 cont.)NPSLESRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIQQGYYGMDVWGQGTTVTVSS 122S-60-15 (SEQ ID NO: 56 cont.)NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTTVTVSS 122S-60-15.1 (SEQ ID NO: 56 cont.)NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTTVTVSS 122S-60-16 (SEQ ID NO: 56 cont.)NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTTVTVSS 122S-60-18 (SEQ ID NO: 56 cont.)NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTTVTVSS 122S-60-19 (SEQ ID NO: 54 cont.)NPSLKSQVTISVDTSKNQFSLELSSVTAADTAVYYCARQGSIQQGYYGMDVWGQGTTVTVSS 122S-60-24 (SEQ ID NO: 56 cont.)NPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMDVWGQGTTVTVSS 122 S-60Alignment ****:*:**************:********************:*******************S-60 HVR H2 (SEQ ID NO: 2 cont.)NPSLKS-------------------------------------------------------- S-60HVR H3 (SEQ ID NO: 6)------------------------------------ARQGSIKQGYYGMDV-----------Sequences of the V_(L) regions of the S-60 antibody variants, alignment to S-60, and HVR locationsS-60 (SEQ ID NO: 79)DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRA 60 S-60-1(SEQ ID NO: 113)DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRA 60 S-60-2(SEQ ID NO: 114)DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRA 60 S-60-3(SEQ ID NO: 115)DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSNGYNYLDWYLQKPGQPPQLLIYLGSNRV 60 S-60-4(SEQ ID NO: 116)DIVMTQSPLSLPVTPGESASISCRSSQGLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNRA 60 S-60-7(SEQ ID NO: 117)DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNRA 60 S-60-8(SEQ ID NO: 118)DIVMTQSPLSLPVTPGGPASISCRSSQSLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNRA 60 S-60-10(SEQ ID NO: 57)DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNRA 60 S-60-11(SEQ ID NO: 58)DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNRA 60 S-60-12(SEQ ID NO: 59)DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSNGYNYLDWYLQKPGQPPQLLIYLGSNRV 60 S-60-13(SEQ ID NO: 57)DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNRA 60 S-60-14(SEQ ID NO: 58)DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNRA 60 S-60-15(SEQ ID NO: 57)DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNRA 60S-60-15.1 (SEQ ID NO: 60)DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSTGYNYLDWYLQKPGQSPQLLIYLGSNRA 60 S-60-16(SEQ ID NO: 77)DIVMTQSPLSLPVTPGEPASISCRSSQSLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNRA 60 S-60-18(SEQ ID NO: 78)DIVMTQSPLSLPVTPGGPASISCRSSQSLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNRA 60 S-60-19(SEQ ID NO: 79)DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRA 60 S-60-24(SEQ ID NO: 80)DIVMTQSPLSLPVTPGESASISCRSSQGLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNRA 60 S-60Alignment ****************  *********.**:**************** ***********.S-60 HVR L1 (SEQ ID NO: 27)------------------------------RSSQSLLHSNGYNYLD-------------- S-60HVR L2 (SEQ ID NO: 29)------------------------------------------------------LGSNRA S-60(SEQ ID NO: 79 cont.)SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112 S-60-1(SEQ ID NO: 113 cont.)SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112 S-60-2(SEQ ID NO: 114 cont.)SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112 S-60-3(SEQ ID NO: 115 cont.)SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112 S-60-4(SEQ ID NO: 116 cont.)SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112 S-60-7(SEQ ID NO: 117 cont.)SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112 S-60-8(SEQ ID NO: 118 cont.)SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112 S-60-10(SEQ ID NO: 57 cont.)SGVPDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGGGTKVEIK 112 S-60-11(SEQ ID NO: 58 cont.)SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQEAPLTFGGGTKVEIK 112 S-60-12(SEQ ID NO: 59 cont.)SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112 S-60-13(SEQ ID NO: 57 cont.)SGVPDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGGGTKVEIK 112 S-60-14(SEQ ID NO: 58 cont.)SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQEAPLTFGGGTKVEIK 112 S-60-15(SEQ ID NO: 57 cont.)SGVPDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGGGTKVEIK 112 S-60-15.1(SEQ ID NO: 60 cont.)SGVPDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGGGTKVEIK 112 S-60-16(SEQ ID NO: 77 cont.)SGVPDRFSGSGSGTDFTLKISRVEAEDVGAYYCMQQQEAPLTFGGGTKVEIK 112 S-60-18(SEQ ID NO: 78 cont.)SGVPDRLSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112 S-60-19(SEQ ID NO: 79 cont.)SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGGGTKVEIK 112 S-60-24(SEQ ID NO: 80 cont.)SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQEAPLTFGGGTKVEIK 112 S-60 Align.******:***************.******.********:************* S-60HVR L2 (SEQ ID NO: 29 cont.)S--------------------------------------------------- S-60HVR L3 (SEQ ID NO: 33)---------------------------------MQQQETPLT----------

TABLE 16 Heavy chain HVR H1 sequences of anti-SORT1 antibodies Ab(s)HVR H1 SEQ ID NO:S-60; S-60-10; 5-60-11; S-60-12; S-60-13; S-60-14; S-60-15 YSISSGYYWG 1[N33 (wt)]; S-60-15.1 [N33T]; S-60-15.2 [N33S]; S-60-15.3[N33G]; S-60-15.4 [N33R]; S-60-15.5 [N33D]; S-60-15.6[N33H]; S-60-15.7 [N33K]; S-60-15.8 [N33Q]; S-60-15.9[N33Y]; S-60-15.10 [N33E]; S-60-15.11 [N33W]; S-60-15.12[N33F]; S-60-15.13 [N33I]; S-60-15.14 [N33V]; S-60-15.15[N33A]; S-60-15.16 [N33M]; S-60-15.17 [N33L]; S-60-16; 5-60-18; S-60-19; S-60-24

TABLE 17 Heavy chain HVR H2 sequences of anti-SORT1 antibodies Ab(s)HVR H2 SEQ ID NO:S-60; S-60-10; S-60-11; S-60-12; S-60-15 [N33 (wt)]; S-60-TIYHSGSTYYNPSL 215.1 [N33T]; S-60-15.2 [N33S]; S-60-15.3 [N33G]; S-60-15.4 KS[N33R]; S-60-15.5 [N33D]; S-60-15.6 [N33H]; S-60-15.7[N33K]; S-60-15.8 [N33Q]; S-60-15.9 [N33Y]; S-60-15.10[N33E]; S-60-15.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13[N33I]; S-60-15.14 [N33V]; S-60-15.15 [N33A]; S-60-15.16[N33M]; S-60-15.17 [N33L]; S-60-16; S-60-18; S-60-19; S-60- 24S-60-13; S-60-14 TIYHSGSTYYNPSL 3 ES Formula I TIYHSGSTYYNPSL 4 X₁SX₁ is K or E

TABLE 18 Heavy chain HVR H3 sequences of anti-SORT1 antibodies Ab(s)HVR H3 SEQ ID NO: S-60-10; S-60-11; S-60-12; S-60-13; S-60-14; S-60-19ARQGSIQQGYYGM 5 DV S-60; S-60-15 [N33 (wt)]; S-60-15.1 [N33T]; S-60-15.2ARQGSIKQGYYGM 6 [N33S]; S-60-15.3 [N33G]; S-60-15.4 [N33R]; S-60-15.5 DV[N33D]; S-60-15.6 [N33H]; S-60-15.7 [N33K]; S-60-15.8[N33Q]; S-60-15.9 [N33Y]; S-60-15.10 [N33E]; S-60-15.11[N33W]; S-60-15.12 [N33F]; S-60-15.13 [N33I]; S-60-15.14[N33V]; S-60-15.15 [N33A]; S-60-15.16 [N33M]; S-60-15.17[N33L]; S-60-16; S-60-18; S-60-24 Formula II ARQGSIX₁QGYYGM 7 DVX₁ is Q or K

TABLE 19 Light chain HVR L1 sequences of anti-SORT1 antibodies Ab(s)HVR L1 SEQ ID NO: S-60-10; S-60-11; S-60-12; S-60-13; S-60-14;RSSQSLLRSNGYNY  8 S-60-15 [N33 (wt)]; S-60-16; S-60-18 LDS-60-15.1 [N33T] RSSQSLLRSTGYNYL  9 D S-60-15.2 [N33S] RSSQSLLRSSGYNYL10 D S-60-15.3 [N33G] RSSQSLLRSGGYNY 11 LD S-60-15.4 [N33R]RSSQSLLRSRGYNY 12 LD S-60-15.5 [N33D] RSSQSLLRSDGYNY 13 LDS-60-15.6 [N33H] RSSQSLLRSHGYNY 14 LD S-60-15.7 [N33K] RSSQSLLRSKGYNY 15LD S-60-15.8 [N33Q] RSSQSLLRSQGYNY 16 LD S-60-15.9 [N33Y] RSSQSLLRSYGYNY17 LD S-60-15.10 [N33E] RSSQSLLRSEGYNYL 18 D S-60-15.11 [N33W]RSSQSLLRSWGYNY 19 LD S-60-15.12 [N33F] RSSQSLLRSFGYNYL 20 DS-60-15.13 [N33I] RSSQSLLRSIGYNYL 21 D S-60-15.14 [N33V] RSSQSLLRSVGYNY22 LD S-60-15.15 [N33A] RSSQSLLRSAGYNY 23 LD S-60-15.16 [N33M]RSSQSLLRSMGYNY 24 LD S-60-15.17 [N33L] RSSQSLLRSLGYNYL 25 DS-60; S-60-19 RSSQSLLHSNGYNY 26 LD S-60-24 RSSQGLLRSNGYNY 27 LDFormula III RSSQX₁LLX₂SX₃GYN 28 YLD X₁ is S or G X₂ is R or HX₃ is N, T, S, G, R, D, H, K, Q, Y, E, W, F, I, V, A, M, or L

TABLE 20 Light chain HVR L2 sequences of anti-SORT1 antibodies Ab(s)HVR L2 SEQ ID NO: S-60; S-60-10; S-60-11; S-60-13; S-60-14; S-60-15LGSNRAS 29 [N33 (wt)]; S-60-15.1 [N33T]; S-60-15.2 [N33S]; S-60-15.3[N33G]; S-60-15.4 [N33R]; S-60-15.5 [N33D]; S-60-15.6[N33H]; S-60-15.7 [N33K]; S-60-15.8 [N33Q]; S-60-15.9[N33Y]; S-60-15.10 [N33E]; S-60-15.11 [N33W]; S-60-15.12[N33F]; S-60-15.13 [N33I]; S-60-15.14 [N33V]; S-60-15.15[N33A]; S-60-15.16 [N33M]; S-60-15.17 [N33L]; S-60-16; S-60-18; S-60-19; S-60-24 S-60-12 LGSNRVS 30 Formula IV LGSNRX₁S 31X₁ is A or V

TABLE 21 Light chain HVR L3 sequences of anti-SORT1 antibodies Ab(s)HVR L3 SEQ ID NO:S-60-10; S-60-11; S-60-13; S-60-14; S-60-15 [N33 (wt)]; S- MQQQEAPLT 3260-15.1 [N33T]; S-60-15.2 [N33S]; S-60-15.3 [N33G]; S-60-15.4 [N33R]; S-60-15.5 [N33D]; S-60-15.6 [N33H]; S-60-15.7[N33K]; S-60-15.8 [N33Q]; S-60-15.9 [N33Y]; S-60-15.10[N33E]; S-60-15.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13[N33I]; S-60-15.14 [N33V]; S-60-15.15 [N33A]; S-60-15.16[N33M]; S-60-15.17 [N33L]; S-60-16; S-60-24S-60; S-60-12; S-60-18; S-60-19 MQQQETPLT 33 Formula V MQQQEX₁PLT 34X₁ is A or T

TABLE 22 Heavy chain framework 1 sequences of anti-SORT1 antibodiesAb(s) VH FR1 SEQ ID NO: S-60-10; S-60-11; S-60-12; S-60-13; S-60-14;S-60-15 [N33 QVQLQESGPGLVKP 35 (wt)]; S-60-15.1 [N33T]; S-60-15.2[N33S]; S-60-15.3 SETLSLTCAVSG [N33G]; S-60-15.4 [N33R]; S-60-15.5[N33D]; S-60-15.6 [N33H]; S-60-15.7 [N33K]; S-60-15.8 [N33Q]; S-60-15.9[N33Y]; S-60-15.10 [N33E]; S-60-15.11 [N33W]; S-60-15.12 [N33F];S-60-15.13 [N33I]; S-60-15.14 [N33V]; S-60-15.15 [N33A]; S-60-15.16[N33M]; S-60-15.17 [N33L]; S-60-16; S- 60-18; S-60-19; S-60-24

TABLE 23 Heavy chain framework 2 sequences of anti-SORT1 antibodiesAb(s) VH FR2 SEQ ID NO: S-60-10; S-60-11; S-60-12; S-60-13; S-60-14;S-60-15 [N33 WIRQPPGKGLEWIG 36 (wt)]; S-60-15.1 [N33T]; S-60-15.2[N33S]; S-60-15.3 [N33G]; S-60-15.4 [N33R]; S-60-15.5 [N33D]; S-60-15.6[N33H]; S-60-15.7 [N33K]; S-60-15.8 [N33Q]; S-60-15.9 [N33Y]; S-60-15.10[N33E]; S-60-15.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13 [N33I];S-60-15.14 [N33V]; S-60-15.15 [N33A]; S-60-15.16 [N33M]; S-60-15.17[N33L]; S-60-16; S- 60-18; S-60-19; S-60-24

TABLE 24 Heavy chain framework 3 sequences of anti-SORT1 antibodiesAb(s) VH FR3 SEQ ID NO: S-60-10; S-60-11; S-60-12; S-60-19QVTISVDTSKNQFSL 37 ELSSVTAADTAVYYC S-60-13; S-60-14; S-60-15 [N33 (wt)];S-60-15.1 [N33T]; S- RVTISVDTSKNQFSL 38 60-15.2 [N33S]; S-60-15.3[N33G]; S-60-15.4 [N33R]; S-60- KLSSVTAADTAVYYC 15.5 [N33D]; S-60-15.6[N33H]; S-60-15.7 [N33K]; S-60- 15.8 [N33Q]; S-60-15.9 [N33Y];S-60-15.10 [N33E]; S-60- 15.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13[N33I]; S-60- 15.14 [N33V]; S-60-15.15 [N33A]; S-60-15.16 [N33M]; S-60-15.17 [N33L]; S-60-16; S-60-18; S-60-24 Formula VI X₁VTISVDTSKNQFS 39LX₂LSSVTAADTAVY YC X₁ is Q or R X₂ is E or K

TABLE 25 Heavy chain framework 4 sequences of anti-SORT1 antibodiesAb(s) VH FR4 SEQ ID NO: S-60-10; S-60-11; S-60-12; S-60-13; S-60-14;S-60-15 [N33 WGQGTTVTVSS 40 (wt)]; S-60-15.1 [N33T]; S-60-15.2 [N33S];S-60-15.3 [N33G]; S-60-15.4 [N33R]; S-60-15.5 [N33D]; S-60-15.6 [N33H];S-60-15.7 [N33K]; S-60-15.8 [N33Q]; S-60-15.9 [N33Y]; S-60-15.10 [N33E];S-60-15.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13 [N33I]; S-60-15.14[N33V]; S-60-15.15 [N33A]; S-60-15.16 [N33M]; S-60-15.17 [N33L];S-60-16; S- 60-18; S-60-19; S-60-24

TABLE 26 Light chain framework 1 sequences of anti-SORT1 antibodiesAb(s) VL FR1 SEQ ID NO: S-60-10; S-60-11; S-60-12; S-60-13; S-60-14;S-60-15 [N33 DIVMTQSPLSLPVTP 41 (wt)]; S-60-15.1 [N33T]; S-60-15.2[N33S]; S-60-15.3 GEPASISC [N33G]; S-60-15.4 [N33R]; S-60-15.5 [N33D];S-60-15.6 [N33H]; S-60-15.7 [N33K]; S-60-15.8 [N33Q]; S-60-15.9 [N33Y];S-60-15.10 [N33E]; S-60-15.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13[N33I]; S-60-15.14 [N33V]; S-60-15.15 [N33A]; S-60-15.16 [N33M];S-60-15.17 [N33L]; S-60-16; S- 60-19 S-60-18 DIVMTQSPLSLPVTP 42 GGPASISCS-60-24 DIVMTQSPLSLPVTP 43 GESASISC Formula VII DIVMTQSPLSLPVTP 44GX₁X₂ASISC X₁ is E or G X₂ is P or S

TABLE 27 Light chain framework 2 sequences of anti-SORT1 antibodiesAb(s) VL FR2 SEQ ID NO: S-60-10; S-60-11; S-60-13; S-60-14; S-60-15 [N33(wt)]; S- WYLQKPGQSPQLLIY 45 60-15.1 [N33T]; S-60-15.2 [N33S]; S-60-15.3[N33G]; S-60- 15.4 [N33R]; S-60-15.5 [N33D]; S-60-15.6 [N33H]; S-60-15.7[N33K]; S-60-15.8 [N33Q]; S-60-15.9 [N33Y]; S-60-15.10 [N33E];S-60-15.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13 [N33I]; S-60-15.14[N33V]; S-60-15.15 [N33A]; S-60-15.16 [N33M]; S-60-15.17 [N33L];S-60-16; S-60-18; S-60-19; S- 60-24 S-60-12 WYLQKPGQPPQLLIY 46 FormulaVIII WYLQKPGQX₁PQLLIY 47 X₁ is S or P

TABLE 28 Light chain framework 3 sequences of anti-SORT1 antibodiesAb(s) VL FR3 SEQ ID NO: S-60-10; S-60-13; S-60-15 [N33 (wt)]; S-60-15.1[N33T]; S- GVPDRFSGSGSGTD 48 60-15.2 [N33S]; S-60-15.3 [N33G]; S-60-15.4[N33R]; S-60- FTLKISRAEAEDVGV 15.5 [N33D]; S-60-15.6 [N33H]; S-60-15.7[N33K]; S-60- YYC 15.8 [N33Q]; S-60-15.9 [N33Y]; S-60-15.10 [N33E];S-60- 15.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13 [N33I]; S-60- 15.14[N33V]; S-60-15.15 [N33A]; S-60-15.16 [N33M]; S-60- 15.17 [N33L]S-60-11; S-60-12; S-60-14; S-60-19; S-60-24 GVPDRFSGSGSGTD 49FTLKISRVEAEDVGV YYC S-60-16 GVPDRFSGSGSGTD 50 FTLKISRVEAEDVGA YYCS-60-18 GVPDRLSGSGSGTD 51 FTLKISRVEAEDVGV YYC Formula IX GVPDRX₁SGSGSGTD52 FTLKISRX₂EAEDVG X₃YYC X₁ is F or L X₂ is A or V X₃ is V or A

TABLE 29 Light chain framework 4 sequences of anti-SORT1 antibodiesAb(s) VL FR4 SEQ ID NO: S-60-10; S-60-11; S-60-12; S-60-13; S-60-14;S-60-15 [N33 FGGGTKVEIK 53 (wt)]; S-60-15.1 [N33T]; S-60-15.2 [N33S];S-60-15.3 [N33G]; S-60-15.4 [N33R]; S-60-15.5 [N33D]; S-60-15.6 [N33H];S-60-15.7 [N33K]; S-60-15.8 [N33Q]; S-60-15.9 [N33Y]; S-60-15.10 [N33E];S-60-15.11 [N33W]; S-60-15.12 [N33F]; S-60-15.13 [N33I]; S-60-15.14[N33V]; S-60-15.15 [N33A]; S-60-15.16 [N33M]; S-60-15.17 [N33L];S-60-16; S- 60-18; S-60-19; S-60-24

TABLE 30 Heavy chain variable region sequences of anti-SORT1 antibodiesAb(s) HCVR SEQ ID NO: S-60-10, S-60-11, S-60-12, S-60-19QVQLQESGPGLVKPSETLSLTCAVSG 54 YSISSGYYWGWIRQPPGKGLEWIGTIYHSGSTYYNPSLKSQVTISVDTSKNQFS LELSSVTAADTAVYYCARQGSIQQGY YGMDVWGQGTTVTVSSS-30-13, S-60-14 QVQLQESGPGLVKPSETLSLTCAVSG 55YSISSGYYWGWIRQPPGKGLEWIGTIY HSGSTYYNPSLESRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQGSIQQGY YGMDVWGQGTTVTVSS S-60, S-60-15 [N33 (wt)],S-60-15.1 QVQLQESGPGLVKPSETLSLTCAVSG 56 [N33T], S-60-15.2 [N33S],S-60-15.3 YSISSGYYWGWIRQPPGKGLEWIGTIY [N33G], S-60-15.4 [N33R],S-60-15.5 HSGSTYYNPSLKSRVTISVDTSKNQFS [N33D], S-60-15.6 [N33H],S-60-15.7 LKLSSVTAADTAVYYCARQGSIKQGY [N33K], S-60-15.8 [N33Q], S-60-15.9YGMDVWGQGTTVTVSS [N33Y], S-60-15.10 [N33E], S-60-15.11 [N33W],S-60-15.12 [N33F], S-60-15.13 [N33I], S-60-15.14 [N33V], S-60-15.15[N33A], S-60-15.16 [N33M], S-60-15.17 [N33L], S-60-16, S-60-18, S-60-24

TABLE 31 Light chain variable region sequences of anti-SORT1 antibodiesAb(s) LCVR SEQ ID NO: S-60-10; S-60-13; S-60-15 [N33 (wt)]DIVMTQSPLSLPVTPGEPASISCRSSQS 57 LLRSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RAEAEDVGVYYCMQQQEAPLTFGGG TKVEIK S-60-11;S-60-14 DIVMTQSPLSLPVTPGEPASISCRSSQS 58 LLRSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RVEAEDVGVYYCMQQQEAPLTFGGG TKVEIK S-60-12DIVMTQSPLSLPVTPGEPASISCRSSQS 59 LLRSNGYNYLDWYLQKPGQPPQLLIYLGSNRVSGVPDRFSGSGSGTDFTLKIS RVEAEDVGVYYCMQQQETPLTFGGG TKVEIK S-60-15.1[N33T] DIVMTQSPLSLPVTPGEPASISCRSSQS 60 LLRSTGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RAEAEDVGVYYCMQQQEAPLTFGGG TKVEIK S-60-15.2[N33S] DIVMTQSPLSLPVTPGEPASISCRSSQS 61 LLRSSGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RAEAEDVGVYYCMQQQEAPLTFGGG TKVEIK S-60-15.3[N33G] DIVMTQSPLSLPVTPGEPASISCRSSQS 62 LLRSGGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RAEAEDVGVYYCMQQQEAPLTFGGG TKVEIK S-60-15.4[N33R] DIVMTQSPLSLPVTPGEPASISCRSSQS 63 LLRSRGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RAEAEDVGVYYCMQQQEAPLTFGGG TKVEIK S-60-15.5[N33D] DIVMTQSPLSLPVTPGEPASISCRSSQS 64 LLRSDGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RAEAEDVGVYYCMQQQEAPLTFGGG TKVEIK S-60-15.6[N33H] DIVMTQSPLSLPVTPGEPASISCRSSQS 65 LLRSHGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RAEAEDVGVYYCMQQQEAPLTFGGG TKVEIK S-60-15.7[N33K] DIVMTQSPLSLPVTPGEPASISCRSSQS 66 LLRSKGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RAEAEDVGVYYCMQQQEAPLTFGGG TKVEIK S-60-15.8[N33Q] DIVMTQSPLSLPVTPGEPASISCRSSQS 67 LLRSQGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RAEAEDVGVYYCMQQQEAPLTFGGG TKVEIK S-60-15.9[N33Y] DIVMTQSPLSLPVTPGEPASISCRSSQS 68 LLRSYGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RAEAEDVGVYYCMQQQEAPLTFGGG TKVEIK S-60-15.10[N33E] DIVMTQSPLSLPVTPGEPASISCRSSQS 69 LLRSEGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RAEAEDVGVYYCMQQQEAPLTFGGG TKVEIK S-60-15.11[N33W] DIVMTQSPLSLPVTPGEPASISCRSSQS 70 LLRSWGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKI SRAEAEDVGVYYCMQQQEAPLTFGG GTKVEIK S-60-15.12[N33F] DIVMTQSPLSLPVTPGEPASISCRSSQS 71 LLRSFGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RAEAEDVGVYYCMQQQEAPLTFGGG TKVEIK S-60-15.13[N33I] DIVMTQSPLSLPVTPGEPASISCRSSQS 72 LLRSIGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RAEAEDVGVYYCMQQQEAPLTFGGG TKVEIK S-60-15.14[N33V] DIVMTQSPLSLPVTPGEPASISCRSSQS 73 LLRSVGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RAEAEDVGVYYCMQQQEAPLTFGGG TKVEIK S-60-15.15[N33A] DIVMTQSPLSLPVTPGEPASISCRSSQS 74 LLRSAGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RAEAEDVGVYYCMQQQEAPLTFGGG TKVEIK S-60-15.16[N33M] DIVMTQSPLSLPVTPGEPASISCRSSQS 75 LLRSMGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RAEAEDVGVYYCMQQQEAPLTFGGG TKVEIK S-60-15.17[N33L] DIVMTQSPLSLPVTPGEPASISCRSSQS 76 LLRSLGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RAEAEDVGVYYCMQQQEAPLTFGGG TKVEIK S-60-16DIVMTQSPLSLPVTPGEPASISCRSSQS 77 LLRSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RVEAEDVGAYYCMQQQEAPLTFGGG TKVEIK S-60-18DIVMTQSPLSLPVTPGGPASISCRSSQS 78 LLRSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRLSGSGSGTDFTLKIS RVEAEDVGVYYCMQQQETPLTFGGG TKVEIK S-60,S-60-19 DIVMTQSPLSLPVTPGEPASISCRSSQS 79 LLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RVEAEDVGVYYCMQQQETPLTFGGG TKVEIK S-60-24DIVMTQSPLSLPVTPGESASISCRSSQG 80 LLRSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKIS RVEAEDVGVYYCMQQQEAPLTFGGG TKVEIK

TABLE 32 Sortilin amino acid sequences SEQ ID Description Sequence NOHuman         10         20         30         40         50 81 SortilinMERPWGAADG LSRWPHGLGL LLLLQLLPPS TLSQDRLDAP PPPAAPLPRW        60         70         80         90        100 SGPIGVSWGLRAAAAGGAFP RGGRWRRSAP GEDEECGRVR DFVAKLANNT       110        120        130        140        150 HQHVFDDLRGSVSLSWVGDS TGVILVLTTF HVPLVIMTFG QSKLYRSEDY       160        170        180        190        200 GKNFKDITDLINNTFIRTEF GMAIGPENSG KVVLTAEVSG GSRGGRIFRS       210        220        230        240        250 SDFAKNFVQTDLPFHPLTQM MYSPQNSDYL LALSTENGLW VSKNFGGKWE       260        270        280        290        300EIHKAVCLAK WGSDNTIFFT TYANGSCKAD LGALELWRTS DLGKSFKTIG       310        320        330        340        350 VKIYSFGLGGRFLFASVMAD KDTTRRIHVS TDQGDTWSMA QLPSVGQEQF       360        370        380        390        400 YSILAANDDMVFMHVDEPGD TGFGTIFTSD DRGIVYSKSL DRHLYTTTGG       410        420        430        440        450 ETDFTNVTSLRGVYITSVLS EDNSIQTMIT FDQGGRWTHL RKPENSECDA       460        470        480        490        500 TAKNKNECSLHIHASYSISQ KLNVPMAPLS EPNAVGIVIA HGSVGDAISV       510        520        530        540        550 MVPDVYISDDGGYSWTKMLE GPHYYTILDS GGIIVAIEHS SRPINVIKFS       560        570        580        590        600 TDEGQCWQTYTFTRDPIYFT GLASEPGARS MNISIWGFTE SFLTSQWVSY       610        620        630        640        650 TIDFKDILERNCEEKDYTIW LAHSTDPEDY EDGCILGYKE QFLRLRKSSV       660        670        680        690        700 CQNGRDYVVTKQPSICLCSL EDFLCDFGYY RPENDSKCVE QPELKGHDLE       710        720        730        740        750 FCLYGREEHLTTNGYRKIPG DKCQGGVNPV REVKDLKKKC TSNFLSPEKQ       760        770        780        790        800 NSKSNSVPIILAIVGLMLVT VVAGVLIVKK YVCGGRFLVH RYSVLQQHAE       810        820        830 ANGVDGVDAL DTASHTNKSG YHDDSDEDLL EMouse Sortilin MERPRGAADG LLRWPLGLLL LLQLLPPAAV GQDRLDAPPP PAPPLLRWAG 82PVGVSWGLRA AAPGGPVPRA GRWRRGAPAE DQDCGRLPDF IAKLTNNTHQ HVFDDLSGSVSLSWVGDSTG VILVLTTFQV PLVIVSFGQS KLYRSEDYGK NFKDITNLIN NTFIRTEFGMAIGPENSGKV ILTAEVSGGS RGGRVFRSSD FAKNFVQTDL PFHPLTQMMY SPQNSDYLLALSTENGLWVS KNFGEKWEEI HKAVCLAKWG PNNIIFFTTH VNGSCKADLG ALELWRTSDLGKTFKTIGVK IYSFGLGGRF LFASVMADKD TTRRIHVSTD QGDTWSMAQL PSVGQEQFYSILAANEDMVF MHVDEPGDTG FGTIFTSDDR GIVYSKSLDR HLYTTTGGET DFTNVTSLRGVYITSTLSED NSIQSMITFD QGGRWEHLRK PENSKCDATA KNKNECSLHI HASYSISQKLNVPMAPLSEP NAVGIVIAHG SVGDAISVMV PDVYISDDGG YSWAKMLEGP HYYTILDSGGIIVAIEHSNR PINVIKFSTD EGQCWQSYVF TQEPIYFTGL ASEPGARSMN ISIWGFTESFITRQWVSYTV DFKDILERNC EEDDYTTWLA HSTDPGDYKD GCILGYKEQF LRLRKSSVCQNGRDYVVAKQ PSVCPCSLED FLCDFGYFRP ENASECVEQP ELKGHELEFC LYGKEEHLTTNGYRKIPGDK CQGGMNPARE VKDLKKKCTS NFLNPTKQNS KSNSVPIILA IVGLMLVTVVAGVLIVKKYV CGGRFLVHRY SVLQQHAEAD GVEALDSTSH AKSGYHDDSD EDLLE RatSortilin MERPRGAADG LLRWPLGLLL LLQLLPPAAV GQDRLDAPPP PAPPLLRWAG 83PVGVSWGLRA AAPGGPVPRA GRWRRGAPAE DQDCGRLPDF IAKLTNNTHQ HVFDDLSGSVSLSWVGDSTG VILVLTTFQV PLVIVSFGQS KLYRSEDYGK NFKDITNLIN NTFIRTEFGMAIGPENSGKV ILTAEVSGGS RGGRVFRSSD FAKNFVQTDL PFHPLTQMMY SPQNSDYLLALSTENGLWVS KNFGEKWEEI HKAVCLAKWG PNNIIFFTTH VNGSCKADLG ALELWRTSDLGKTFKTIGVK IYSFGLGGRF LFASVMADKD TTRRIHVSTD QGDTWSMAQL PSVGQEQFYSILAANDDMVF MHVDEPGDTG FGTIFTSDDR GIVYSKSLDR HLYTTTGGET DFTNVTSLRGVYITSTLSED NSIQSMITFD QGGRWEHLQK PENSKCDATA KNKNECSLHI HASYSISQKLNVPMAPLSEP NAVGIVIAHG SVGDAISVMV PDVYISDDGG YSWAKMLEGP HYYTILDSGGIIVAIEHSNR PINVIKFSTD EGQCWQSYVF SQEPVYFTGL ASEPGARSMN ISIWGFTESFLTRQWVSYTI DFKDILERNC EENDYTTWLA HSTDPGDYKD GCILGYKEQF LRLRKSSVCQNGRDYVVAKQ PSICPCSLED FLCDFGYFRP ENASECVEQP ELKGHELEFC LYGKEEHLTTNGYRKIPGDR CQGGMNPARE VKDLKKKCTS NFLNPKKQNS KSSSVPIILA IVGLMLVTVVAGVLIVKKYV CGGRFLVHRY SVLQQHAEAD GVEALDTASH AKSGYHDDSD EDLLE

TABLE 33 S-60-15 peptide sequences SEQ ID Description Sequence NOAsp-box motif (S/T)-X-(D/N)-X-X-X-X-(W/F/Y) 84 X is any amino acidAsp-box motif X-X-(S/T)-X-(D/N)-X-G-X-(T/S)-(W/F/Y)-X 85 X is any aminoacid Asp-box motif SSDFAKNF 86 in human Sortilin (residues 200-207)S-60-15 NQFSLK 87 Peptide S-60-15 Q GYYG M DVWG Q GTTVTVSSASTK 88Peptide S-60-15 DYFPEPVTVS WN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTK 89Peptide S-60-15 DTL M ISR 90 Peptide S-60-15TPEVTCVVVDVSQEDPEVQFNWYVDGVEVH N AK 91 Peptide S-60-15 VVSVLTVLHQDWLN GK 92 Peptide S-60-15 EPQVYTLPPSQEE M TK 93 Peptide S-60-15 N QVSLTCLVK94 Peptide S-60-15 GFYPSDIAVEWES N GQPENNYK 95 Peptide S-60-15WQEGNVFSCSV M HEALH N HYTQK 96 Peptide S-60-15 DIV MTQSPLSLPVTPGEPASISCR 97 Peptide S-60-15 SS Q SLLR 98 Peptide S-60-15 S NGYNYLDWYLQKPGQSPQLLIYLGSNR 99 Peptide S-60-15 AEAEDVGVYYC MQQQEAPLTFGGGTK 100 Peptide S-60-15 SGTASVVCLL N NFYPR 101 PeptideS-60-15 VD N ALQSGNSQESVTEQDSK 102 Peptide

TABLE 34 Fc domain amino acid sequences SEQ ID Description Sequence NOhuIgG1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT 103 LALAPS FcSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQK SLSLSPGK huIgG1ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT 104 LALAPS FcSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKV without C-DKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPE terminal lysineVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQK SLSLSPG

TABLE 35 Full-length heavy chain amino acid sequences SEQ ID DescriptionSequence NO S-60-10, S-60-11, S-60-12, S-QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGW 105 60-19 with Fc LALAPS withIRQPPGKGLEWIGTIYHSGSTYYNPSLKSQVTISVDTS C-terminal LysineKNQFSLELSSVTAADTAVYYCARQGSIQQGYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK S-60-10, S-60-11,S-60-12, S- QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGW 106 60-19 with FcLALAPS IRQPPGKGLEWIGTIYHSGSTYYNPSLKSQVTISVDTS without C-terminal LysineKNQFSLELSSVTAADTAVYYCARQGSIQQGYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPG S-60-13, S-60-14 with FcQVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGW 135 LALAPS with C-terminalIRQPPGKGLEWIGTIYHSGSTYYNPSLESRVTISVDTSK LysineNQFSLKLSSVTAADTAVYYCARQGSIQQGYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK S-60-13, S-60-14 withFc QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGW 136 LALAPS without C-IRQPPGKGLEWIGTIYHSGSTYYNPSLESRVTISVDTSK terminal LysineNQFSLKLSSVTAADTAVYYCARQGSIQQGYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPG S-60, S-60-15 [N33(wt)], S- QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGW 137 60-15.1 [N33T],S-60-15.2 IRQPPGKGLEWIGTIYHSGSTYYNPSLKSRVTISVDTS [N33S], S-60-15.3[N33G], KNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMD S-60-15.4 [N33R], S-60-15.5VWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAAL [N33D], S-60-15.6 [N33H],GCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL S-60-15.7 [N33K], S-60-15.8YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVE [N33Q], S-60-15.9 [N33Y],PKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMIS S-60-15.10 [N33E], S-60-RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT 15.11 [N33W], S-60-15.12KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS [N33F], S-60-15.13 [N33I],NKALPASIEKTISKAKGQPREPQVYTLPPSRDELTKNQ S-60-15.14 [N33V], S-60-VSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD 15.15 [N33A], S-60-15.16SDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH [N33M], S-60-15.17 [N33L],YTQKSLSLSPGK S-60-16, S-60-18, S-60-24 with Fc LALAPS with C- terminalLysine S-60, S-60-15 [N33 (wt)], S-QVQLQESGPGLVKPSETLSLTCAVSGYSISSGYYWGW 138 60-15.1 [N33T], S-60-15.2IRQPPGKGLEWIGTIYHSGSTYYNPSLKSRVTISVDTS [N33S], S-60-15.3 [N33G],KNQFSLKLSSVTAADTAVYYCARQGSIKQGYYGMD S-60-15.4 [N33R], S-60-15.5VWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAAL [N33D], S-60-15.6 [N33H],GCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL S-60-15.7 [N33K], S-60-15.8YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVE [N33Q], S-60-15.9 [N33Y],PKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMIS S-60-15.10 [N33E], S-60-RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT 15.11 [N33W], S-60-15.12KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS [N33F], S-60-15.13 [N33I],NKALPASIEKTISKAKGQPREPQVYTLPPSRDELTKNQ S-60-15.14 [N33V], S-60-VSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD 15.15 [N33A], S-60-15.16SDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH [N33M], S-60-15.17 [N33L],YTQKSLSLSPG S-60-16, S-60-18, S-60-24 with Fc LALAPS without C-terminalLysine

TABLE 36 Full-length light chain amino acid sequences SEQ DescriptionSequence ID NO S-60-10; S-60-13; S- DIVMTQSPLSLPVTPGEPASIS 139 60-15[N33 (wt)] CRSSQSLLRSNGYNYLDWYL QKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRAEA EDVGVYYCMQQQEAPLTFGG GTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE AKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSS PVTKSFNRGEC S-60-11; S-60-14 DIVMTQSPLSLPVTPGEPASIS140 CRSSQSLLRSNGYNYLDWYL QKPGQSPQLLIYLGSNRASGV PDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQEAPLTFGG GTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-12 DIVMTQSPLSLPVTPGEPASIS 141 CRSSQSLLRSNGYNYLDWYLQKPGQPPQLLIYLGSNRVSGV PDRFSGSGSGTDFTLKISRVEA EDVGVYYCMQQQETPLTFGGGTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPRE AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSS PVTKSFNRGEC S-60-15.1 [N33T]DIVMTQSPLSLPVTPGEPASIS 142 CRSSQSLLRSTGYNYLDWYL QKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRAEA EDVGVYYCMQQQEAPLTFGG GTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE AKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSS PVTKSFNRGEC S-60-15.2 [N33S] DIVMTQSPLSLPVTPGEPASIS143 CRSSQSLLRSSGYNYLDWYL QKPGQSPQLLIYLGSNRASGV PDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGG GTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-15.3 [N33G] DIVMTQSPLSLPVTPGEPASIS 144CRSSQSLLRSGGYNYLDWYL QKPGQSPQLLIYLGSNRASGV PDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGG GTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-15.4 [N33R] DIVMTQSPLSLPVTPGEPASIS 145CRSSQSLLRSRGYNYLDWYL QKPGQSPQLLIYLGSNRASGV PDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGG GTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-15.5 [N33D] DIVMTQSPLSLPVTPGEPASIS 146CRSSQSLLRSDGYNYLDWYL QKPGQSPQLLIYLGSNRASGV PDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGG GTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-15.6 [N33H] DIVMTQSPLSLPVTPGEPASIS 119CRSSQSLLRSHGYNYLDWYL QKPGQSPQLLIYLGSNRASGV PDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGG GTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-15.7 [N33K] DIVMTQSPLSLPVTPGEPASIS 120CRSSQSLLRSKGYNYLDWYL QKPGQSPQLLIYLGSNRASGV PDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGG GTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-15.8 [N33Q] DIVMTQSPLSLPVTPGEPASIS 121CRSSQSLLRSQGYNYLDWYL QKPGQSPQLLIYLGSNRASGV PDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGG GTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-15.9 [N33Y] DIVMTQSPLSLPVTPGEPASIS 122CRSSQSLLRSYGYNYLDWYL QKPGQSPQLLIYLGSNRASGV PDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGG GTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-15.10 [N33E] DIVMTQSPLSLPVTPGEPASIS 123CRSSQSLLRSEGYNYLDWYL QKPGQSPQLLIYLGSNRASGV PDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGG GTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-15.11 [N33W] DIVMTQSPLSLPVTPGEPASIS 124CRSSQSLLRSWGYNYLDWYL QKPGQSPQLLIYLGSNRASGV PDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGG GTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-15.12 [N33F] DIVMTQSPLSLPVTPGEPASIS 125CRSSQSLLRSFGYNYLDWYL QKPGQSPQLLIYLGSNRASGV PDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGG GTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-15.13 [N33I] DIVMTQSPLSLPVTPGEPASIS 126CRSSQSLLRSIGYNYLDWYLQ KPGQSPQLLIYLGSNRASGVP DRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGGG TKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-15.14 [N33V] DIVMTQSPLSLPVTPGEPASIS 127CRSSQSLLRSVGYNYLDWYL QKPGQSPQLLIYLGSNRASGV PDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGG GTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-15.15 [N33A] DIVMTQSPLSLPVTPGEPASIS 128CRSSQSLLRSAGYNYLDWYL QKPGQSPQLLIYLGSNRASGV PDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGG GTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-15.16 [N33M] DIVMTQSPLSLPVTPGEPASIS 129CRSSQSLLRSMGYNYLDWYL QKPGQSPQLLIYLGSNRASGV PDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGG GTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-15.17 [N33L] DIVMTQSPLSLPVTPGEPASIS 130CRSSQSLLRSLGYNYLDWYL QKPGQSPQLLIYLGSNRASGV PDRFSGSGSGTDFTLKISRAEAEDVGVYYCMQQQEAPLTFGG GTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-16 DIVMTQSPLSLPVTPGEPASIS 131 CRSSQSLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGV PDRFSGSGSGTDFTLKISRVEA EDVGAYYCMQQQEAPLTFGGGTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPRE AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSS PVTKSFNRGEC S-60-18DIVMTQSPLSLPVTPGGPASIS 132 CRSSQSLLRSNGYNYLDWYL QKPGQSPQLLIYLGSNRASGVPDRLSGSGSGTDFTLKISRVEA EDVGVYYCMQQQETPLTFGG GTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE AKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSS PVTKSFNRGEC S-60, S-60-19 DIVMTQSPLSLPVTPGEPASIS133 CRSSQSLLHSNGYNYLDWYL QKPGQSPQLLIYLGSNRASGV PDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQQQETPLTFGG GTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC S-60-24 DIVMTQSPLSLPVTPGESASIS 134 CRSSQGLLRSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGV PDRFSGSGSGTDFTLKISRVEA EDVGVYYCMQQQEAPLTFGGGTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPRE AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSS PVTKSFNRGEC

1. An antibody that binds to a Sortilin protein, wherein the antibodycomprises a heavy chain variable region and a light chain variableregion, wherein (a) the heavy chain variable region comprises an HVR-H1comprising the amino acid sequence YSISSGYYWG (SEQ ID NO: 1), an HVR-H2comprising the amino acid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), anHVR-H3 comprising the amino acid sequence ARQGSIQQGYYGMDV (SEQ ID NO:5); and the light chain variable region comprises an HVR-L1 comprisingthe amino acid sequence RSSQSLLRSNGYNYLD (SEQ ID NO: 8), an HVR-L2comprising the amino acid sequence LGSNRAS (SEQ ID NO: 29), and anHVR-L3 comprising the amino acid sequence MQQQEAPLT (SEQ ID NO: 32); (b)the heavy chain variable region comprises an HVR-H1 comprising the aminoacid sequence YSISSGYYWG (SEQ ID NO: 1), an HVR-H2 comprising the aminoacid sequence TIYHSGSTYYNPSLKS (SEQ ID NO: 2), an HVR-H3 comprising theamino acid sequence ARQGSIQQGYYGMDV (SEQ ID NO: 5); and the light chainvariable region comprises an HVR-L1 comprising the amino acid sequenceRSSQSLLRSNGYNYLD (SEQ ID NO: 8), an HVR-L2 comprising the amino acidsequence LGSNRVS (SEQ ID NO: 30), and an HVR-L3 comprising the aminoacid sequence MQQQETPLT (SEQ ID NO: 33); (c) the heavy chain variableregion comprises an HVR-H1 comprising the amino acid sequence YSISSGYYWG(SEQ ID NO: 1), an HVR-H2 comprising the amino acid sequenceTIYHSGSTYYNPSLES (SEQ ID NO: 3), an HVR-H3 comprising the amino acidsequence ARQGSIQQGYYGMDV (SEQ ID NO: 5); and the light chain variableregion comprises an HVR-L1 comprising the amino acid sequenceRSSQSLLRSNGYNYLD (SEQ ID NO: 8), an HVR-L2 comprising the amino acidsequence LGSNRAS (SEQ ID NO: 29), and an HVR-L3 comprising the aminoacid sequence MQQQEAPLT (SEQ ID NO: 32); (d) the heavy chain variableregion comprises an HVR-H1 comprising the amino acid sequence YSISSGYYWG(SEQ ID NO: 1), an HVR-H2 comprising the amino acid sequenceTIYHSGSTYYNPSLKS (SEQ ID NO: 2), an HVR-H3 comprising the amino acidsequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and the light chain variableregion comprises an HVR-L1 comprising the amino acid sequenceRSSQSLLRSNGYNYLD (SEQ ID NO: 8), an HVR-L2 comprising the amino acidsequence LGSNRAS (SEQ ID NO: 29), and an HVR-L3 comprising the aminoacid sequence MQQQEAPLT (SEQ ID NO: 32); (e) the heavy chain variableregion comprises an HVR-H1 comprising the amino acid sequence YSISSGYYWG(SEQ ID NO: 1), an HVR-H2 comprising the amino acid sequenceTIYHSGSTYYNPSLKS (SEQ ID NO: 2), an HVR-H3 comprising the amino acidsequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and the light chain variableregion comprises an HVR-L1 comprising the amino acid sequenceRSSQSLLRSTGYNYLD (SEQ ID NO: 9), an HVR-L2 comprising the amino acidsequence LGSNRAS (SEQ ID NO: 29), and an HVR-L3 comprising the aminoacid sequence MQQQEAPLT (SEQ ID NO: 32); (f) the heavy chain variableregion comprises an HVR-H1 comprising the amino acid sequence YSISSGYYWG(SEQ ID NO: 1), an HVR-H2 comprising the amino acid sequenceTIYHSGSTYYNPSLKS (SEQ ID NO: 2), an HVR-H3 comprising the amino acidsequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and the light chain variableregion comprises an HVR-L1 comprising the amino acid sequenceRSSQSLLRSNGYNYLD (SEQ ID NO: 8), an HVR-L2 comprising the amino acidsequence LGSNRAS (SEQ ID NO: 29), and an HVR-L3 comprising the aminoacid sequence MQQQETPLT (SEQ ID NO: 33); (g) the heavy chain variableregion comprises an HVR-H1 comprising the amino acid sequence YSISSGYYWG(SEQ ID NO: 1), an HVR-H2 comprising the amino acid sequenceTIYHSGSTYYNPSLKS (SEQ ID NO: 2), an HVR-H3 comprising the amino acidsequence ARQGSIQQGYYGMDV (SEQ ID NO: 5); and the light chain variableregion comprises an HVR-L1 comprising the amino acid sequenceRSSQSLLHSNGYNYLD (SEQ ID NO: 26), an HVR-L2 comprising the amino acidsequence LGSNRAS (SEQ ID NO: 29), and an HVR-L3 comprising the aminoacid sequence MQQQETPLT (SEQ ID NO: 33); (h) the heavy chain variableregion comprises an HVR-H1 comprising the amino acid sequence YSISSGYYWG(SEQ ID NO: 1), an HVR-H2 comprising the amino acid sequenceTIYHSGSTYYNPSLKS (SEQ ID NO: 2), an HVR-H3 comprising the amino acidsequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and the light chain variableregion comprises an HVR-L1 comprising the amino acid sequenceRSSQGLLRSNGYNYLD (SEQ ID NO: 27), an HVR-L2 comprising the amino acidsequence LGSNRAS (SEQ ID NO: 29), and an HVR-L3 comprising the aminoacid sequence MQQQEAPLT (SEQ ID NO: 32); (i) the heavy chain variableregion comprises an HVR-H1 comprising the amino acid sequence YSISSGYYWG(SEQ ID NO: 1), an HVR-H2 comprising the amino acid sequenceTIYHSGSTYYNPSLKS (SEQ ID NO: 2), an HVR-H3 comprising the amino acidsequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and the light chain variableregion comprises an HVR-L1 comprising the amino acid sequenceRSSQSLLRSNGYNYLD (SEQ ID NO: 10), an HVR-L2 comprising the amino acidsequence LGSNRAS (SEQ ID NO: 29), and an HVR-L3 comprising the aminoacid sequence MQQQEAPLT (SEQ ID NO: 32); or (j) the heavy chain variableregion comprises an HVR-H1 comprising the amino acid sequence YSISSGYYWG(SEQ ID NO: 1), an HVR-H2 comprising the amino acid sequenceTIYHSGSTYYNPSLKS (SEQ ID NO: 2), an HVR-H3 comprising the amino acidsequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and the light chain variableregion comprises an HVR-L1 comprising the amino acid sequenceRSSQSLLRSNGYNYLD (SEQ ID NO: 21), an HVR-L2 comprising the amino acidsequence LGSNRAS (SEQ ID NO: 29), and an HVR-L3 comprising the aminoacid sequence MQQQEAPLT (SEQ ID NO: 32).
 2. The antibody of claim 1,wherein the anti-Sortilin antibody: (a) decreases cell surface levels ofSortilin more than the level of decrease caused by an anti-Sortilinantibody comprising a heavy chain variable region comprising thesequence of SEQ ID NO: 56 and a light chain variable region comprisingthe sequence of SEQ ID NO: 79; (b) increases extracellular levels ofProgranulin more than the level of increase caused by an anti-Sortilinantibody comprising a heavy chain variable region comprising thesequence of SEQ ID NO: 56 and a light chain variable region comprisingthe sequence of SEQ ID NO: 79; (c) inhibits the interaction betweenSortilin and Progranulin more than the level of inhibition caused by ananti-Sortilin antibody comprising a heavy chain variable regioncomprising the sequence of SEQ ID NO: 56 and a light chain variableregion comprising the sequence of SEQ ID NO: 79; or (d) any combinationof (a)-(c).
 3. The antibody of claim 1, wherein the antibody has adissociation constant (K_(D)) for human Sortilin that is at least1.1-fold lower than an anti-Sortilin antibody comprising a heavy chainvariable region comprising the sequence of SEQ ID NO: 56 and a lightchain variable region comprising the sequence of SEQ ID NO: 79, whereinthe K_(D) is determined by FACS.
 4. The antibody of claim 1, wherein theantibody has a dissociation constant (K_(D)) for human Sortilin thatranges from about 5.0E-10 M to about 1.0E-9 M wherein the K_(D) isdetermined by FACS, or about 250-500 pM wherein the K_(D) is determinedby Bio-layer interferometry.
 5. An antibody that binds to a Sortilinprotein, wherein the antibody comprises a heavy chain variable regionand a light chain variable region, wherein: the heavy chain variableregion comprises an HVR-H1 comprising the amino acid sequence YSISSGYYWG(SEQ ID NO: 1), an HVR-H2 comprising the amino acid sequenceTIYHSGSTYYNPSLKS (SEQ ID NO: 2), and an HVR-H3 comprising the amino acidsequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and the light chain variableregion comprises an HVR-L1 comprising the amino acid sequenceRSSQSLLRSTGYNYLD (SEQ ID NO: 9), an HVR-L2 comprising the amino acidsequence LGSNRAS (SEQ ID NO: 29), and an HVR-L3 comprising the aminoacid sequence MQQQEAPLT (SEQ ID NO: 32).
 6. An antibody that binds to aSortilin protein, wherein the antibody comprises a heavy chain variableregion and a light chain variable region, wherein: the heavy chainvariable region comprises an HVR-H1 comprising the amino acid sequenceYSISSGYYWG (SEQ ID NO: 1), an HVR-H2 comprising the amino acid sequenceTIYHSGSTYYNPSLKS (SEQ ID NO: 2), and an HVR-H3 comprising the amino acidsequence ARQGSIKQGYYGMDV (SEQ ID NO: 6); and the light chain variableregion comprises an HVR-L1 comprising the amino acid sequenceRSSQSLLRSNGYNYLD (SEQ ID NO: 8), an HVR-L2 comprising the amino acidsequence LGSNRAS (SEQ ID NO: 29), and an HVR-L3 comprising the aminoacid sequence MQQQEAPLT (SEQ ID NO: 32).
 7. An antibody that binds to aSortilin protein, wherein the antibody comprises: (a) a heavy chainvariable region comprising the amino acid sequence of SEQ ID NO: 54, anda light chain variable region comprising the amino acid sequence of SEQID NO: 57; (b) a heavy chain variable region comprising the amino acidsequence of SEQ ID NO: 54, and a light chain variable region comprisingthe amino acid sequence of SEQ ID NO: 58; (c) a heavy chain variableregion comprising the amino acid sequence of SEQ ID NO: 54, and a lightchain variable region comprising the amino acid sequence of SEQ ID NO:59; (d) a heavy chain variable region comprising the amino acid sequenceof SEQ ID NO: 55, and a light chain variable region comprising the aminoacid sequence of SEQ ID NO: 57; (e) a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO: 55, and a light chainvariable region comprising the amino acid sequence of SEQ ID NO: 58; (f)a heavy chain variable region comprising the amino acid sequence of SEQID NO: 56, and a light chain variable region comprising the amino acidsequence of SEQ ID NO: 57; (g) a heavy chain variable region comprisingthe amino acid sequence of SEQ ID NO: 56, and a light chain variableregion comprising the amino acid sequence of SEQ ID NO: 77; (h) a heavychain variable region comprising the amino acid sequence of SEQ ID NO:56, and a light chain variable region comprising the amino acid sequenceof SEQ ID NO: 78; (i) a heavy chain variable region comprising the aminoacid sequence of SEQ ID NO: 54, and a light chain variable regioncomprising the amino acid sequence of SEQ ID NO: 79; (j) a heavy chainvariable region comprising the amino acid sequence of SEQ ID NO: 56, anda light chain variable region comprising the amino acid sequence of SEQID NO: 80; (k) a heavy chain variable region comprising the amino acidsequence of SEQ ID NO: 56, and a light chain variable region comprisingthe amino acid sequence of SEQ ID NO: 60; (l) a heavy chain variableregion comprising the amino acid sequence of SEQ ID NO: 56, and a lightchain variable region comprising the amino acid sequence of SEQ ID NO:61; or (m) a heavy chain variable region comprising the amino acidsequence of SEQ ID NO: 56, and a light chain variable region comprisingthe amino acid sequence of SEQ ID NO:
 72. 8. An antibody that binds to aSortilin protein, wherein the antibody comprises a heavy chain variableregion comprising the amino acid sequence of SEQ ID NO: 56 and a lightchain variable region comprising the amino acid sequence of SEQ ID NO:60.
 9. An antibody that binds to a Sortilin protein, wherein theantibody comprises a heavy chain variable region comprising the aminoacid sequence of SEQ ID NO: 56 and a light chain variable regioncomprising the amino acid sequence of SEQ ID NO:
 57. 10. An antibodythat binds to a Sortilin protein, wherein the antibody comprises a heavychain comprising the amino acid sequence of SEQ ID NO: 137 and a lightchain comprising the amino acid sequence of SEQ ID NO:
 142. 11. Anantibody that binds to a Sortilin protein, wherein the antibodycomprises a heavy chain comprising the amino acid sequence of SEQ ID NO:138 and a light chain comprising the amino acid sequence of SEQ ID NO:142.
 12. The antibody of claim 1 or claim 7, wherein the antibody is ofthe IgG class, the IgM class, or the IgA class.
 13. The antibody ofclaim 12, wherein the antibody is of the IgG class and has an IgG1,IgG2, IgG3, or IgG4 isotype.
 14. The antibody of claim 13, wherein: (a)the antibody is an IgG1 or IgG2 isotype and the Fc region comprises anamino acid substitution at position P331S, wherein the numbering of theresidue position is according to EU numbering; (b) the antibody is anIgG1 isotype and the Fc region comprises amino acid substitutions atpositions L234A, L235A, and P331S, wherein the numbering of the residueposition is according to EU numbering; (c) the antibody is an IgG1,IgG2, or IgG4 isotype and the Fc region comprises an amino acidsubstitution at position N297A, wherein the numbering of the residueposition is according to EU numbering; or (d) the Fc region comprises anamino acid substitution at positions S267E and L328F, wherein thenumbering of the residue position is according to EU numbering.
 15. Theantibody of claim 14, wherein the antibody is an IgG1 isotype and the Fcregion comprises amino acid substitutions at positions L234A, L235A, andP331S, wherein the numbering of the residue position is according to EUnumbering.
 16. The antibody of claim 1, wherein the Sortilin protein isa human Sortilin protein.
 17. The antibody of claim 16, wherein thehuman Sortilin protein comprises the amino acid sequence of SEQ ID NO:81.
 18. The antibody of claim 1, wherein the Sortilin protein is awild-type protein, a naturally occurring variant, or a disease variant.19. The antibody of claim 1, wherein the antibody is a human antibody, abispecific antibody, a monoclonal antibody, a multivalent antibody, or aconjugated antibody.
 20. The antibody of claim 19, wherein the antibodyis a bispecific antibody recognizing a first antigen and a secondantigen.
 21. The antibody of claim 20, wherein the first antigen isSortilin and the second antigen is an antigen facilitating transportacross the blood-brain-barrier.
 22. The antibody of claim 21, whereinthe second antigen is selected from the group consisting of Sortilin,transferrin receptor (TR), insulin receptor (HIR), insulin-like growthfactor receptor (IGFR), low-density lipoprotein receptor relatedproteins 1 and 2 (LPR-1 and 2), diphtheria toxin receptor, CRM197, allama single domain antibody, TMEM 30(A), a protein transduction domain,TAT, Syn-B, penetratin, a poly-arginine peptide, an angiopep peptide,basigin, Glut1, CD98hc, and ANG1005.
 23. The antibody of claim 1,wherein the antibody is an antibody fragment that binds to a humanSortilin protein.
 24. The antibody of claim 23, wherein the antibodyfragment is an Fab, Fab′, Fab′-SH, F(ab′)2, Fv, or scFv fragment.
 25. Anisolated nucleic acid comprising a nucleic acid sequence encoding theantibody of claim
 1. 26. A vector comprising the nucleic acid of claim25.
 27. An isolated host cell comprising the vector of claim
 26. 28. Amethod of producing an antibody that binds to Sortilin, comprisingculturing the host cell of claim 27 so that the antibody is produced.29. (canceled)
 30. An isolated antibody that binds to Sortilin producedby the method of claim
 28. 31. A pharmaceutical composition comprisingthe antibody of any one of claim 1, 7, 10, or 11 and a pharmaceuticallyacceptable carrier.
 32. A method of preventing, reducing risk for, ortreating an individual having a disease, disorder, or injury, comprisingadministering to the individual a therapeutically effective amount ofthe antibody of claim
 1. 33-34. (canceled)
 35. A method of inhibitingone or more of neuroinflammation, axonopathy characterized by shortaxonal outgrowth and aberrant branching, microglial activation, andinflammatory response in an individual in need thereof, comprisingadministering to the individual a therapeutically effective amount ofthe anti-Sortilin antibody of claim
 1. 36. A method of promoting one ormore of wound healing, autophagy, and clearance of aggregate proteins inan individual in need thereof, comprising administering to theindividual a therapeutically effective amount of the anti-Sortilinantibody of claim
 1. 37. (canceled)
 38. A method of decreasingexpression of one or more pro-inflammatory mediators in an individual inneed thereof, comprising administering to the individual atherapeutically effective amount of the anti-Sortilin antibody ofclaim
 1. 39. (canceled)
 40. A method of inhibiting the interactionbetween Sortilin and Progranulin, the method comprising exposing a cellexpressing Sortilin to the anti-Sortilin antibody of claim
 1. 41-44.(canceled)
 45. A method of increasing levels of Progranulin in anindividual in need thereof, the method comprising administering to theindividual a therapeutically effective amount of the anti-Sortilinantibody of claim
 1. 46-47. (canceled)
 48. A method of decreasing levelsof Sortilin in an individual in need thereof, the method comprisingadministering to the individual a therapeutically effective amount ofthe anti-Sortilin antibody of claim
 1. 49-53. (canceled)